TK8 and Allegro™ Single-Use Systems and Manufacturing

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Qualification Package

TK8 and Allegro™ Single-Use Systems and Manufacturing Process Comparison Document Number: USTR3382

Revision Number: 1.0

Date: 25th March 2020

Author: C. Piton

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Contents

1 Introduction ................................................................................................................................... 4 2 Replacement of TK8 Film Based Products Strategy .................................................................... 4

2.1 Product Definition ............................................................................................................. 5 2.2 Replacement Products ..................................................................................................... 6 2.3 TK8 Film Based Products and Allegro Film Based Products Comparison ...................... 7 2.4 TK8 and Allegro Film-Based Products Manufacturing Approach .................................. 12

3 Qualification Data and Comparison Between Allegro Film and TK8 Film .................................. 13 3.1 Physical Descriptions ..................................................................................................... 13 3.1.1 Film Compositions.......................................................................................................... 13 3.1.2 Cosmetic Aspect ............................................................................................................ 14 3.1.3 Tack ............................................................................................................................... 14 3.1.4 Handling Recommendations .......................................................................................... 14 3.1.5 Summary ........................................................................................................................ 15 3.2 Physical Characteristics of Allegro and TK8 Films ........................................................ 15 3.2.1 Tensile Strength at Break .............................................................................................. 15 3.2.2 Elongation at Break ........................................................................................................ 15 3.2.3 Elastic Modulus (Young Modulus) ................................................................................. 16 3.2.4 Puncture Resistance ...................................................................................................... 16 3.2.5 Dart Impact ..................................................................................................................... 16 3.2.6 Flex Crack Resistance ................................................................................................... 16 3.2.7 Tear Resistance ............................................................................................................. 17 3.3 Summary of Comparison Between Allegro Film and TK8 Film ..................................... 17 3.4 Physical Characteristics of Allegro Film from 2 Lots ...................................................... 18 3.5 Allegro Film Shelf Life Data ........................................................................................... 19 3.5.1 Results Discussion ......................................................................................................... 20 3.5.2 Summary ........................................................................................................................ 24 3.6 Barrier Properties ........................................................................................................... 24 3.6.1 Test Data ........................................................................................................................ 24 3.6.2 Comparison Between Films ........................................................................................... 25 3.6.3 Summary ........................................................................................................................ 25 3.7 Compliance Standards on Raw Materials and Film ....................................................... 25 3.8 Extractables Studies on TK8 and Allegro Films ............................................................. 26 3.8.1 Extraction Conditions ..................................................................................................... 26 3.8.2 Analytical Methods ......................................................................................................... 28 3.8.3 ICH Q3D Elemental Impurities ....................................................................................... 28 3.8.4 Summary of ICH Q3C Residual Solvents and ICH Q3D Elemental Impurities ............. 30 3.8.5 Summary ........................................................................................................................ 30 3.9 Chemical Compatibility .................................................................................................. 30

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4 Manufacturing Process Change Qualification ............................................................................ 31 4.1 Manufacturing Environment ........................................................................................... 32 4.2 Manufacturing Processes .............................................................................................. 32 4.2.1 Differences in the Film Format ....................................................................................... 34 4.2.2 Leak Testing Differences ............................................................................................... 34 4.2.3 Summary ........................................................................................................................ 35 4.3 SU systems Qualifications: New Cleanroom ................................................................. 35 4.3.1 SU systems and Components Qualifications ................................................................. 36 4.3.2 Standards for SU Systems Containing Chambers ......................................................... 37 4.3.3 In-Process Controls ....................................................................................................... 37 4.3.4 Summary ........................................................................................................................ 38

5 Data Supporting Applications: Performances Assessment ........................................................ 38 5.1 Mixing Applications ........................................................................................................ 38 5.1.1 Equivalence of Designs ................................................................................................. 38 5.1.2 Durability Tests .............................................................................................................. 38 5.2 Shipping and Transportation .......................................................................................... 39 5.2.1 Intra-Site Transport Verification ..................................................................................... 39 5.2.2 Shipping Lab Simulations .............................................................................................. 39 5.3 Freezing Applications ..................................................................................................... 40 5.3.1 Preliminary Study – TK8 Film ........................................................................................ 40 5.3.2 Preliminary Study – Allegro Film .................................................................................... 40 5.3.3 Conclusions .................................................................................................................... 40

6 Conclusions ................................................................................................................................ 41 Appendix 1. List of Standards ............................................................................................................... 42 Appendix 2. Comparison of Physical Characteristics Results for TK8 and Allegro Film ...................... 45

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1 Introduction

This document provides qualification information about the characteristics and manufacturing processes currently available to support the transition from the discontinued Pall TK8 film to the Pall Allegro film.

Characterization data had been previously generated for both films at the time of the initial qualification. As the films originate from two different companies, test results were generated by different laboratories, following different test standards and/or using different test conditions.

To allow a more comprehensive comparison, Pall Biotech has run a test campaign to generate new datasets on both films under identical test conditions, especially for physical characteristics, gas barrier properties and shelf life supporting data.

To support film transition, this comparison report includes key characterization data, with results from a new dataset (e.g. for physical characteristics) along with results issued from the initial qualification dataset. After comparative analysis, recommendations are provided to address the change from TK8 to Allegro film.

2 Replacement of TK8 Film Based Products Strategy

The TK8 film is used for the manufacturing of the single-use (SU) products from different families.

Table 1 Product families impacted by the discontinuation of TK8 film

Product Family Consumable Type Volumes Impacted

Associated Hardware Impact

Notification Letter

Storage

2D shape biocontainer

50 mL, 100 mL, 150 mL, 250 mL, 500 mL, 1 L, 2 L, 5 L, 10 L, 20 L, 50 L

No impact on Allegro plastic trays CN-RPII-B2QK6N


50 L, 100 L, 200 L, 500 L, 1000 L, 2000 L, 2500 L, 3000 L

No impact on standard Allegro stainless steel or plastic hardware CN-RPII-B2QK6N

Mixing

LevMixer® system, round & cubical shape

30 L, 50 L, 100 L, 200 L, 350 L, 400 L, 425 L, 500 L, 560 L, 650 L, 1000 L, 2000 L, 3000 L

No impact on standard LevMixer hardware CN-RPII-B2QK6N

Magnetic Mixer system, round & cubical shape

30 L, 50 L, 100 L, 200 L, 350 L, 400 L, 425 L, 500 L, 650 L, 1000 L, 1500 L, 1900 L, 2000 L, 3000 L

No impact on standard Magnetic Mixer hardware CN-RPII-B2QK6N

Wand mixer system 5 L, 10 L, 20 L, 50 L, 100 L, 200 L

No impact on standard Wand mixer hardware CN-RPII-B2QK6N

PadMixer® system 1 25 L, 50 L, 200 L, 500 L, 1000 L

PadMixer hardware will be discontinued (separate notification to be released) CN-RPII-B2QK6N

Jet mixer system 50 L, 200 L, 500 L, 1000 L Discontinued hardware CN-RPII-B2QK6N


Table 1 continued

Powder handling bags – Powder transfer vessel (PTV)2

PTV, 2 in. and 4 in. 150 mL, 2.5 L, 5 L, 15 L, 30 L, 100 L

Not applicable – No dedicated PTV hardware CN-RPII-B4BPF9

Liner, 2 in. and 4 in. 5 L Not applicable CN-RPII-B4BPF9

PCB, powder handling chutes bag, 4 in. Not applicable Not applicable CN-RPII-B4BPF9

Bioreactors

iCELLis® foam bag 5 L No impact on iCELLis hardware CN-RPII-B2QK6N

PadReactor® system 25 L, 50 L, 125 L, 250 L, 600 L, 1200 L Discontinued hardware CN-RPII-B4BJGR

PadReactor Mini bioreactor 16 L Discontinued hardware CN-RPII-B4BJGR

1 PadMixer hardware will be discontinued and therefore not available for new application.

2 Powder handling bags do not exist with Allegro film but can be replaced by existing products made with PD2 film. These products are qualified, and a validation guide is available (USTR3092a). For further information, please contact your local Pall Biotech specialist.

2.1 Product Definition Pall Biotech is manufacturing single-use (SU) assembled systems which include Pall own and outsourced components. The single-use assembled systems are also defined as SU systems in this document and represent products provided to end-users.

1. Standard SU systems are approved by Pall Biotech product management. All end-users are using the same part numbers.

2. Customized SU systems are approved by one end user. One part number is specific to a given end-user.

Figure 1 Definition of standard vs customized SU components and SU systems

A standard chamber is a bag with a defined number of ports, sizes and locations (top, bottom, front, sides). Pall Biotech is establishing a list of standard chambers with different types of impeller for mixing chambers or without impeller for storage chambers. As part of a continuous quality improvement initiative, Pall has decided to move away from customizing chamber designs and to focus on standardization to develop a more sustainable manufacturing process, mitigating potential risks, to ensure continuity of supply. Consequently, the Allegro film-based products, replacing TK8 film-based products, may show differences on chamber designs without impacting fitment into standard mixing or storage hardware.

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2.2 Replacement Products For each of the product families, standard Allegro film-based chambers are qualified to be used either in standard Allegro SU system or in customized Allegro SU systems.

For storage applications, the Allegro product family has been available for more than 10 years and fits into standard Allegro storage plastic or stainless-steel totes and trays. For the replacement of some custom 3D TK8 film-based chamber designs it may be required to change hardware in order to use the standard Allegro film-based chambers.

For mixing applications, all Allegro film-based chambers were created as part of the TK8 film discontinuation project, to fit into standard hardware for LevMixer, Magnetic Mixer, Wand mixer and PadMixer systems.

Table 2 Available replacement products made with Allegro film

Product Family

Consumable Product Type Technology Available

Equivalent Standard Allegro Available 1

Alternate Allegro product 2

Storage


Historical standard Pall Allegro storage biocontainers

50 mL, 100 mL, 250 mL, 500 mL, 1 L, 2 L, 5 L, 10 L, 20 L, 50 L

150 mL – Use 100 mL or 250 mL


Historical standard Pall Allegro storage biocontainers

50 L, 100 L, 200 L, 500 L, 1000 L, 1500 L, 2000 L, 3000 L 2500 L – Use 3000 L

Mixing

LevMixer system, round & cubical shape

Standard Allegro chambers fitting into standard LevMixer hardware

30 L, 50 L, 100 L, 200 L, 350 L, 400 L, 500 L, 650 L, 1000 L, 2000 L

425 L – Use 500 L 560 L – Use 650 L 3000 L – Use Magnetic Mixer system

Magnetic Mixer system, round & cubical shape

Standard Allegro chambers fitting into standard Magnetic Mixer hardware

30 L, 50 L, 100 L, 200 L, 350 L, 400 L, 500 L, 650 L, 1000 L, 1500 L, 2000 L, 3000 L

425 L – Use 500 L 1900 L – Use 2000 L

Wand mixer system

Standard Allegro chambers fitting into standard Wand mixer hardware

5 L, 10 L, 20 L, 50 L, 100 L, 200 L None

PadMixer system

Allegro chambers fitting into existing PadMixer hardware

25 L, 50 L, 200 L, 500 L, 1000 L

Allegro chambers only for the replacement in existing applications. For new application, consider alternate mixing technology.

Jet mixer system

Standard Allegro chambers fitting into standard LevMixer or Magnetic Mixer hardware None

Move to another technology - LevMixer or Magnetic Mixer system

Bioreactors

iCELLis foam bags

Allegro foam bags fitting iCELLis technology 5 L None

PadReactor system, 25 L to 1200 L

Standard Allegro chambers fitting standard STR hardware None

Move to another technology – Allegro STR bioreactors

PadReactor Mini bioreactor, 16 L to 50 L

1 Available Allegro equivalent standard volumes.

2 Alternative Allegro products for missing volumes.


For each standard SU system manufactured with TK8 film, the corresponding item manufactured with Allegro film has been created with a new part number for storage and mixing standard systems. The range of chamber volumes covered by standard systems remains the same:

• 50 mL to 50 L for 2D storage

• 50 L to 3000 L for 3D storage

• 30 L to 2000 L for mixing round tanks

• 50 L to 2000 L for mixing cubical tanks

In case of questions regarding your standard system design conversion, please contact your Pall representative.

2.3 TK8 Film Based Products and Allegro Film Based Products Comparison The SU system design will be reproduced when transitioning from TK8 film to Allegro film, while TK8 film-based storage and mixing chambers will be replaced by standard Allegro film-based chambers.

Tables 3 - 8 below show the main characteristics of standard Allegro film-based chambers which may differ from existing TK8 film-based chambers. For LevMixer, Magnetic Mixer and Wand mixer chambers the mixing element will not change and its dimensions, shape, material and welding remain the same. For PadMixer chamber, the mixing element (pad) is protected by a sleeve made of TK8 film product contact layer which is changed for an Allegro film based sleeve. The pad dimensions, shape, material and welding remain the same.

At the time of the qualification package writing, the PadMixer Allegro film-based chambers are not created. Equivalent PadMixer chamber designs to existing TK8 film-based chambers will be created after completion of durability testing expected end April 2020.

Table 3 Characteristics of the 2D Allegro film-based storage chambers

Chamber Volume 50 mL, 100 mL, 250 mL, 500 mL, 1 L, 2 L 5 L, 10 L, 20 L, 50 L

Configuration 2D rectangle 2D rectangle

Dimensions Standard Allegro Standard Allegro

Fitment Boat, 3 hose barb ports Boat, 4 hose barb ports

Fitment location Bottom Bottom

Fitment material HDPE HDPE

HDPE: High density polyethylene

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Table 4 Characteristics of the 3D Allegro film storage biocontainer designs

Chamber Volume 50 L, 100 L, 200 L, 500 L, 1000 L, 1500 L 1000 L, 2000 L, 3000 L

Configuration 3D rectangle 3D rectangle (modular tote shape)

Dimensions Standard Allegro Standard Allegro

Top fitment Plate, 2 inlet + 1 sampling hose barb ports None

Bottom fitment Single, 1 outlet, hose barb port + 1 sampling hose barb port (50 L only) Single, 3 inlet/outlet TC ports

Fitment material HDPE HDPE

HDPE: High density polyethylene TC: Tri-clover♦


Table 5 Characteristics of the Allegro film Wand mixer chambers

Chamber Volume 5 L 10 L 20 L 50 L 100 L 200 L

Powder Port (Top) Single TC, 2 in. Single TC, 2 in. Single TC, 2 in. Single TC, 2 in. Single TC, 4 in. Single TC, 4 in.

Top Fitment Single ¼ in. + ⅜ in. Single ¼ in. + ⅜ in. or port plate ½ in., ½ in., ¼ in.

Single ¼ in. + ⅜ in. or port plate ½ in., ½ in., ¼ in.

Single ⅜ in. + ⅜ in. or port plate ½ in., ½ in., ¼ in.

Single ½ in. + ⅜ in. or port plate ½ in., ½ in., ¼ in.

Single ½ in. + ⅜ in. or port plate ½ in., ½ in., ¼ in.

Side Fitment (Front) Single ⅜ in. Single ⅜ in. or ½ in.

Port plate ½ in., ½ in., ¼ in. or ¾ in., ¾ in., ¼ in. or single ⅜ in.

Port plate ½ in., ½ in., ¼ in. or ¾ in., ¾ in., ¼ in.



Bottom Fitment None None None Pall drain valve, ½ in.

Pall drain valve, ½ in. or easy drain valve, ½ in.

Pall drain valve, ½ in. or easy drain valve, ½ in.

Fitment Material HDPE HDPE HDPE HDPE LDPE + PSU

LDPE + PSU HDPE

LDPE + PSU HDPE

HDPE: High Density Polyethylene; TC: Tri-clover; PSU: Polysulfone; LDPE: low-density polyethylene


Table 6 Characteristics of the Allegro film cubical LevMixer and Magnetic Mixer chambers

Chamber Volume 50 L 100 L 200 L 400 L 650 L 1000 L 1500 L 2000 L 3000 L

Powder Port (Top)

Single TC, 4 in.

Single TC, 4 in.

Single TC, 4 in.

Single TC, 4 in.

Single TC, 4 in.

Single TC, 4 in.

Single TC, 4 in.

Single TC, 4 in.

Single TC, 4 in.

Top Fitment

Single ¼ in. + ¼ in. and/or port plate ½ in., ½ in., ¼ in.

Single ¼ in. + ¼ in. and/or port plate ½ in., ½ in., ¼ in.

Single ¼ in. + ¼ in. and port plate ½ in., ½ in., ¼ in. or 1 in., ½ in., ¼ in. or ¾ in., ½ in., ¼ in.




Single ¼ in. + ¼ in. and port plate ¾ in., ½ in., ¼ in.

Single ¼ in. + ¼ in. and port plate ¼ in., ½ in., 1 in. or ¼ in., ½ in., ¾ in.

Single ¼ in. + ¼ in. and port plate 1 in., ½ in., ¼ in.

Side Fitment (Front)

Port plate ¾ in., ¾ in., ½ in. or ¾ in., ¾ in., ¼ in.

Port plate ¾ in., ¾ in., ½ in. or ¾ in., ¾ in., ¼ in.

Port plate ¾ in., ¾ in., ½ in.




Single ¾ in. + ½ in. + ¼ in.

Single ¾ in. + ¾ in. + ½ in.

Single ¾ in. + ¾ in. + ½ in.

Bottom Fitment Single ½ in.

Pall drain valve, ½ in.

Pall drain valve, ½ in. + single ¼ in. or single 1 in. + ¼ in. or single ¾ in. + ¼ in.




Single ¾ in. + ¼ in.

Single ¾ in. + ¼ in. or single 1 in. + ¼ in.

Single 1 in. + ¼ in.

Fitment Material HDPE

LDPE + PSU HDPE

LDPE + PSU HDPE

LDPE + PSU HDPE

LDPE + PSU HDPE

LDPE + PSU HDPE HDPE HDPE HDPE

HDPE: High density polyethylene TC: Tri-clover PSU: Polysulfone PBT: Polybutylene terephthalate


Table 7 Characteristics of the Allegro film round LevMixer and Magnetic Mixer chambers

Chamber Volume 30 L 50 L 100 L 200 L 350 L 500 L 1000 L 2000 L

Powder Port (Top)

Single TC, 2 in.

Single TC, 4 in.

Single TC, 4 in.

Single TC, 4 in.

Single TC, 4 in.

Single TC, 4 in.

Single TC, 4 in.

Single TC, 4 in.

Top Fitment

Single ¼ in. + ¼ in. and port plate ½ in. ½ in. ¼ in. or ¾ in. ¾ in. ½ in.

Single ¼ in. + ¼ in. and/or port plate ½ in. ½ in. ¼ in.

Single ¼ in. + ¼ in. and/or port plate ½ in. ½ in. ¼ in.

Single ¼ in. + ¼ in. and port plate ½ in. ½ in. ¼ in. or 1 in. ½ in. ¼ in. or ¾ in. ½ in. ¼ in.

Single ¼ in. + ¼ in. and port plate ½ in. ½ in. ¼ in.

Single ¼ in. + ¼ in. and port plate ¾ in. ½ in. ¼ in. or ½ in. ½ in. ¼ in. or ¼ in. ½ in. 1 in.

Single ¼ in. + ¼ in. and/or port plate ¼ in. 1 in. 1 in. or ¼ in. ¾ in. ¾ in. or ½ in. ½ in. ¼ in.

Single ¼ in. + ¼ in. and port plate 1 in. ½ in. ¼ in. or ¾ in. ½ in. ¼ in.

Side Fitment (Front)

Port plate ¾ in. ¾ in. ½ in. or ½ in. ½ in. ¼ in.

Port plate ¾ in. ¾ in. ½ in. and single ¼ in.

Port plate ¾ in. ¾ in. ½ in. or ¾ in. ¾ in. ¼ in.


None or single ¾ in. ¾ in. ½ in. or single ¾ in. ¾ in. ¼ in.


Port plate ¾ in. + ¾ in. + ½ in.


Bottom Fitment None Single, ½ in.

Pall drain valve, ½ in.

Pall drain valve, ½ in. or single ¾ in. or 1 in.

None or Pall drain valve, ½ in.

Pall drain valve, ½ in. or single ¾ in. or 1 in.

Pall drain valve ½ in. or Pall drain valve ¾ in. or 1 in.

Pall drain valve ¾ in. or 1 in.

Fitment Material HDPE HDPE

LDPE + PSU HDPE

LDPE + PSU HDPE

LDPE + PSU HDPE

LDPE + PSU HDPE

LDPE + PSU PBT HDPE

PBT HDPE

HDPE: High density polyethylene TC: Tri-clover PSU: Polysulfone PBT: Polybutylene terephthalate

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Table 8 Characteristics of the Allegro film iCELLis foam bag

Chamber volume 5 L

Configuration 3D rectangle

Dimensions 164 mm x 224 mm x 142 mm

Lifting hole Lip design

Top fitment type Single, 1 hose barb port, ½ in.

Side fitment type Single, 1 hose barb port, ⅜ in. Bottom fitment type Single, 1 hose barb port, ½ in.

Fitment material HDPE

HDPE: High Density Polyethylene

All Allegro film-based chambers fit into standard hardware and it is recommended to check configuration of current hardware in use to confirm fitment compatibility with standard Allegro film-based chambers.

For any questions regarding your chamber design conversion, please contact your Pall representative.

2.4 TK8 and Allegro Film-Based Products Manufacturing Approach Historically, TK8 film-based products were manufactured and assembled (SU Systems) at the Pall Life Sciences BVBA manufacturing site in Hoegaarden, Belgium. With the discontinuation of the TK8 film, a new cleanroom has now been qualified in this Hoegaarden Belgium facility, to extend manufacturing capabilities using Allegro film. The qualification of this additional facility will ensure continuity in the supply chain.

Once the manufacturing process fully validated, the approach for Allegro bag chamber families will be a primary manufacturing site and a contingency site, as described in Table 9, in the event of disruption or major manufacturing delay from the primary site. Currently, the Pall Life Sciences Puerto Rico LLC manufacturing site in Fajardo, Puerto Rico is not qualified for the manufacturing of SU chambers.

Table 9 Pall manufacturing sites locations for Allegro film bag chambers

Allegro Film Products Primary Manufacturing Site Back-Up Manufacturing Site

Allegro 2D storage biocontainers Medemblik, The Netherlands None

Allegro 3D storage biocontainers Medemblik, The Netherlands Hoegaarden, Belgium

Allegro LevMixer chambers Hoegaarden, Belgium Medemblik, The Netherlands

Allegro Magnetic Mixer chambers Hoegaarden, Belgium Medemblik, The Netherlands

Allegro Wand mixer chambers Hoegaarden, Belgium Medemblik, The Netherlands

Allegro PadMixer chambers Hoegaarden, Belgium None

iCELLis bioreactor with Allegro foam bags Hoegaarden, Belgium None


The approach for SU systems assembly will not change and can be performed from one of the three qualified Pall Biotech manufacturing sites:

• Pall Life Sciences Belgium BVBA in Hoegaarden, Belgium

• Pall Medistad BV in Medemblik, The Netherlands

• Pall Life Sciences Puerto Rico LLC in Fajardo, Puerto Rico

3 Qualification Data and Comparison Between Allegro Film and TK8 Film

Both films belong to Pall's portfolio of SU technologies. To ensure a clear distinction, they will be named “TK8" and "Allegro" in this document, even if TK8 film is often named as "Allegro TK8" in Pall's technical documents.

TK8 film was originally designed by ATMI LifeSciences and has been marketed since 2009. When Pall acquired ATMI in 2014 the TK8 film was incorporated into Pall's portfolio of SU technologies.

In comparison, Allegro film has been marketed since the end of 2007 and originally used by Pall in the manufacture their SU systems.

3.1 Physical Descriptions 3.1.1 Film Compositions The Allegro film is a co-extruded film, comprising an inert ultra-low-density polyethylene (ULDPE) inner layer, an ethylene vinyl alcohol (EVOH) gas barrier middle layer and a low-density polyethylene (LDPE) outer layer. This simple structure results in a very low level of extractables associated with equivalent barrier properties.

The TK8 film is constructed from laminated layers of polyamide (PA), EVOH and ULDPE (Figure 2). The outer PA layer provides robust puncture resistance, strength, and excellent thermal stability. The EVOH layer is the gas barrier layer. The ULDPE layers provide flexibility, integrity and an ultra-clean, ultra-pure, low-extractables product contacting layer. The inner, fluid-contact, ULDPE layer used in the TK8 film is blow-extruded in-house by Pall under cleanroom conditions (0.2 μm filtered air). Lamination is also performed under controlled, ultra-clean conditions.

For both films the product contact layer is ultra-pure ULDPE. It should be noted that the ULDPE used in TK8 film and in Allegro film are different material grades, coming from different manufacturers.

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Figure 2 Layer structure of films

3.1.2 Cosmetic Aspect Allegro film is clearer, if only slightly, and more transparent than TK8 film. TK8 film has also a marginally darker, brownish tone, due to its nylon outer layer.

Typical marks appearing on the film due to folding are different between TK8 and Allegro films, also attributable to their different structural composition.

3.1.3 Tack Tack refers to the tendency of a film to stick, or adhere, to a surface. Bag chambers manufactured with Allegro film are a little tackier than those manufactured with TK8 film, as the Allegro outer LDPE layer is a stickier material than the nylon outer layer of TK8 film.

3.1.4 Handling Recommendations Because of the tackier outer layer of the Allegro film, the unpacking of these systems may require additional attention, as components such as tubing, may tend to adhere more to the bag chamber.

It is worth noting that the higher tack of the Allegro film could affect set-up and self-deployment of the 3D bag chambers, but this is not considered to pose a significant risk. Allegro 3D bag chambers have a reliable usability, as demonstrated by 10 years of experience without issues reported by end-users. Deployment tests performed by Pall on Allegro systems (for mixing and storage) showed that limited manual manipulation is needed to guarantee good deployment.

It is therefore recommended that operators take specific care of set-up and deployment when introducing the first systems in Allegro film, to identify any difference versus the TK8 version. Transitioning from systems manufactured with TK8 film to systems manufactured with Allegro film may require updating manual handling procedures.

Please refer to USD 25231 ‘Instruction for Use for Allegro 3D Biocontainers’ for further handling information or for further assistance please contact your technical support engineer.

1 USD 2523c, Instructions For Use - Allegro 3D Biocontainers 100, 200, 500, 1000 and 1500 L


3.1.5 Summary Transitioning from systems manufactured with TK8 film to systems manufactured with Allegro film may require updating manual handling procedures.

3.2 Physical Characteristics of Allegro and TK8 Films The following section, and data, directly compare the physical characteristics of both films, using the same tests that were included in the original validation guide of the TK8 film. Puncture resistance, tear resistance, flex crack resistance and dart impact were not part of the original qualification package of the Allegro film and have been generated to allow a more complete comparison with TK8 film. This comparative data can be used as an input when performing a risk assessment when transitioning from TK8 to Allegro film.

The physical characteristics test data were measured during a test campaign in the second half of 2018, on samples gamma-irradiated at a 50 kGy dose. Table 31 in Appendix 2 summarizes all physical characteristics results for TK8 and Allegro films.

When applicable, the measurements have been done on film samples in 2 different directions:

• Machine direction (MD) – corresponds to the direction of winding of a roll of film

• Transverse direction (TD) - corresponds to the direction of the axis of rotation of a roll of film

Comparing data in these 2 directions allows an evaluation of the film's level of anisotropy. Both films show low level of anisotropy, as expected from the Allegro film and the TK8 nylon layer manufacturing processes.

Table 10 Thickness for Allegro film and TK8 film

Physical Characteristics Allegro Film TK8 Film

Thickness 0.325 mm 0.250 mm

3.2.1 Tensile Strength at Break This test measures the force required to stretch the film to its breaking point. Reported result is the average of 5 samples. Sample strips are 15 mm wide and tensile force is applied at an elongation rate of 200 mm/min.

Table 11 Tensile strength at break for Allegro film and TK8 film

Direction Allegro Film TK8 Film

Machine direction 15 MPa 31 MPa

Transverse direction 13 MPa 34 MPa

3.2.2 Elongation at Break This test measures the elongation of the film to its breaking point. Reported result is the average of 5 samples. Sample strips are 15 mm wide and tensile force is applied at an elongation rate of 200 mm/min.

Table 12 Elongation at break for Allegro film and TK8 film


Machine direction 526% 133%

Transverse direction 450% 102%

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3.2.3 Elastic Modulus (Young Modulus) This test measures the resistance of the film to elastic deformation. Reported result is the average of 5 samples. It is calculated using least-squares fit on the initial linear portion of the stress-strain curve of the film. Sample strips are 15 mm wide and tensile force is applied at an elongation rate of 200 mm/min.

Table 13 Elastic modulus (Young modulus) for Allegro film and TK8 film


Machine direction 293 MPa 486 MPa

Transverse direction 295 MPa 478 MPa

3.2.4 Puncture Resistance Puncture resistance measures the load required to puncture the film with a defined tool (maximum load) and the penetration distance (deflection) at which the film actually breaks. Reported result is the average of 10 samples.

Table 14 Puncture resistance for Allegro film and TK8 film

Measurements Allegro Film TK8 Film

Maximum load 103 N 150 N

Deflection at maximum load 20 mm 13 mm

3.2.5 Dart Impact This test measures the resistance of the film to the impact of a free-falling dart, dropped on a specimen clamped in a test fixture. It establishes the weight of the dart when 50 % of the specimens fail under the conditions identified. Two methods (A and B) are prescribed in ASTM D1709-01, corresponding each to a specific dart size, drop height and range of weights that are added to the dart. Method A corresponds to the smallest drop height, dart size and weight range. Method B is used for the TK8 film. For the Allegro film, a hybrid approach had to be used: method B drop height and dart size had to be used, with weight range of method A. Indeed, the Allegro film resists to method A with its highest weight but breaks when using method B with its lowest weight. Using same dart size and height allows a straightforward comparison between TK8 and Allegro films. Due to the complexity of the test, 1 sample only is tested.

Table 15 Dart impact for Allegro film and TK8 film

Physical Characteristics Allegro Film TK8 Film

Dart impact 193 g 1105 g

3.2.6 Flex Crack Resistance This test evaluates the flex crack resistance of the film, by quantifying the formation of pinholes in a sealed tube film sample subjected to cycles combining twisting and horizontal folding. Quantity of pinholes are reported after 500, 1000, and 5000 cycles. Due to the complexity of the test, 2 samples are tested for each condition. Average value is reported in Table 16.


Table 16 Flex crack resistance in machine direction (MD) and transverse direction (TD) for Allegro film and TK8 film

Number of Cycles

Allegro Film TK8 Film

MD TD MD TD

500 cycles 0 pinholes 0 pinholes 0 pinholes 0 pinholes

1000 cycles 0 pinholes 0 pinholes 0 pinholes 0 pinholes

5000 cycles 5.5 pinholes 5.5 pinholes 6 pinholes 4.5 pinholes

Figure 3 Flex crack resistance test sample

Weld

As shown in Figure 3 the weld is parallel to the machine direction for the MD sample, and parallel to the transverse direction for the TD sample.

3.2.7 Tear Resistance This test is measuring the force needed to propagate tearing. The average force used from 25% to 75% of the tearing distance is reported. Reported result is the average of 5 samples.

Table 17 Tear resistance in machine direction (MD) for Allegro film and TK8 film


Machine direction 8.6 N 15 N

Transverse direction 12 N 22 N

3.3 Summary of Comparison Between Allegro Film and TK8 Film Film physical characteristics confirm what can be deducted from the construction and materials: the nylon layer dictates most of the TK8 film physical characteristics. It adds stiffness, which is demonstrated by higher elastic modulus and higher tensile strength, while elongation at break is lower. This means that TK8 film is physically more robust than Allegro film, but the more flexible Allegro film will stretch more before rupture. This is also confirmed with puncture and impact resistance tests, TK8 film showing higher dart impact resistance and puncture resistance, but lower deflection before puncture.

For the Allegro film, the dart impact test must be operated outside the parameters prescribed by the standard: a mix of methods A and B must be used as explained above. It provides a clear comparison with TK8 film, but cannot be used to compare different batches of Allegro film, and has therefore not been used in shelf life study. Pall considers that higher dart impact resistance of the TK8 film does not represent a significant difference in normal use conditions: in any case, the bag chambers must be protected from potential external aggression by their bag chamber holder. As shown by shipping and transportation simulations (see Section 5.2), an Allegro film bag chamber can withstand significant shocks when properly placed and maintained it its holder.

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Flex crack resistance is similar between both films. A good resistance is observed up to 1000 cycles, representing a very harsh challenge, and pinholes are systematically observed after 5000 cycles. As already presented in original TK8 film validation guide, total absence of pinholes cannot be guaranteed after 500 or 1000 cycles (specification is < 2.0 pinholes and < 4.0 pinholes on average, respectively after 500 cycles and 1000 cycles), and high variability is observed on results after 5000 cycles.

Film physical characterization data is mostly used as reference data, to verify that a defined bias (e.g. aging, irradiation, exposure to chemicals) is not having a detrimental impact on the film. It is not possible to directly deduct from this data if a film is adequate for a defined bioprocess application. This must be demonstrated by verification tests mimicking more closely the intended application (see Section 5 for durability tests for mixing or shipping and transport simulations). With this perspective, Pall does not consider one film being superior to the other. Allegro film has been used successfully for the manufacturing of SU chambers since 2007.

3.4 Physical Characteristics of Allegro Film from 2 Lots A comparison was made between 2 lots of Allegro films manufactured in 2017 over the course of 1 week. One of those lots (lot 1) was used in a previous section to provide comparison data versus TK8 film. This section provides a lot-to-lot comparison between 2 Allegro film lots. Samples included in the table below were gamma-irradiated at a 50 kGy dose. The same quantity of samples as described in Section 3.2 has been used for each timepoint and average values are reported.

Table 18 Physical characteristics comparison – 2 lots of Allegro film

Physical Characteristics Direction Allegro Film – Lot 1 Allegro Film – Lot 2

Tensile Strength at Break MD 15 MPa 15 MPa

TD 13 MPa 14 MPa

Elongation at Break MD 526% 512%

TD 450% 519%

Elastic Modulus (Young Modulus) MD 293 MPa 313 MPa

TD 295 MPa 301 MPa

Puncture Resistance

+ Maximum load 103 N 108 N

+ Deflection at maximum load 20 mm 20 mm

Dart Impact 193 g 290 g

Flex Crack Resistance

+ 500 cycles

MD 0 pinholes 0 pinholes

TD 0 pinholes 0 pinholes

+ 1000 cycles

MD 0 pinholes 0 pinholes


+ 5000 cycles

MD 5.5 pinholes 8.5 pinholes

TD 5.5 pinholes 9 pinholes

Tear Resistance MD 8.6 N 7.1 N

TD 12 N 20 N

Differences in tensile strength, elongation and elastic modulus are aligned with the typical relative standard deviation (RSD) observed between samples of the same lot (10%).

Same observation applies to the puncture resistance results, the maximum load showing similar RSD while the deflection at maximum load shows higher variability (RSD up to 15%).


The high variability in dart impact test results confirms the inadequacy of this test to follow Allegro film performance, as explained above.

Variations in flex crack resistance results are observed between the 2 sub-lots. As explained above, these variations are typical of this test. Variability observed here agrees with previous experience on TK8 film (average pinholes up to 8.7 on samples gamma-irradiated at 50 kGy).

High variability is observed for the tear resistance test, with an RSD between samples of the same lot around 30%.

3.5 Allegro Film Shelf Life Data Gamma-irradiated samples, at a 50 kGy dose, have been exposed to accelerated aging at 55°C. Equivalent aging time at 25 °C has been calculated according ASTM F1980-16, with a Q10 aging factor of 2.

Following table summarizes the measurements at different timepoints, covering Pall's claim of 2 years shelf life after gamma irradiation for the Allegro film. The same quantity of samples as described in Section 3.2 has been used for each timepoint and average values are reported.

Table 19 Shelf life for 2 different lots of Allegro film

Physical Characteristics Direction Allegro Film Data

Lot Number Lot 1 Lot 2

Time Points T0 T8.5M T17M T25M T0 T8.5M T17M T25M

Tensile Strength at Break [MPa]

MD 15 13 14 14 15 13 14 13

TD 13 13 14 14 14 14 13 13

Elongation at Break [%]

MD 526 431 479 438 512 398 445 377

TD 450 380 485 444 519 525 453 397

Elastic (Young) Modulus [MPa]

MD 293 288 280 302 313 290 303 313

TD 295 258 295 289 301 288 297 275

Puncture Resistance

+ Maximum load [N] 103 107 103 103 108 105 104 103

+ Deflection at maximum load [mm] 20 16 15 14 20 14 15 14

Tear Resistance – Average Force [N]

MD 9 14 7 19 7 13 15 26

TD 12 17 16 17 20 12 16 26

Flex Crack Resistance [# of Pinholes]

+ 500 cycles

MD 0 0 0 0 0 0 0 0

TD 0 0 0 0 0 0 0 0

+ 1000 cycles

MD 0 0.5 0 1.0 0 1.0 0.5 0.5

TD 0 0 0.5 0.5 0 0 0.5 1.0

+ 5000 cycles

MD 5.5 5.0 4.5 8.0 8.5 8.5 9.5 9.5

TD 5.5 5.0 6.0 9.5 9.0 9.5 7.5 8.5

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3.5.1 Results Discussion Results at each time point have been put in graphs, Figure 4 through Figure 9, for each physical characteristic and each lot, to make the review easier. Confidence interval at 95% is used to illustrate the variability for each time point.

3.5.1.1 Tensile Strength No significant change is observed during the 25 months of aging.

The lack of overlap between confidence intervals at T0 and at T8.5M for tensile strength at break - MD is not considered as technically significant, considering the typical manufacturing and measurement variabilities.

Figure 4 Tensile strength at break

3.5.1.2 Elongation at Break Results on lot 2 might suggest that a minor decrease in elongation at break is appearing after aging. This trend is however not confirmed on lot 1, and lower values observed on last time points of lots 2 must be balanced with the typical, well-known, variability of this test (e.g. T8.5M of lot 1 in TD is of lower value than T25M of lot 2 in TD) – see Figure 5.

Deciding with certainty between these two options is not possible. However, this does not change the typical behavior of the film, which remains ductile, with high elongation at break, constantly above 350%. It is these characteristics which are important.

It can therefore be concluded that there is no significant change in elongation at break during the 25 months of aging.


Figure 5 Elongation at break

3.5.1.3 Elastic Modulus No significant change is observed during the 25 months of aging.

Higher variability than usual is observed for Lot 2 - TD at 25 months (and to a lesser extend for Lot 1 - TD at 8.5 months). This might be linked to the limited quantity of samples used to calculate the time point average value.

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Figure 6 Elastic (Young) modulus

3.5.1.4 Puncture Resistance No significant change observed on maximum load for puncture resistance between the 2 lots during the 25 months of aging.

Figure 7 Puncture resistance - maximum load

A decrease is observed in deflection at puncture, mainly between T0 and T8.5M – Figure 8. This suggests some loss in film's resilience, more visible on this characteristic than on other tests like puncture-maximum load or elongation at break.

Such a decrease is consistent with observations on TK8 (up to 20% decrease between point corresponding to T0 and T17M for TK8 film, according to the original TK8 validation guide USTR3008), with a higher magnitude with Allegro film (-25% on average, between T0 and T17M).


Deflection at maximum load remains significantly higher for Allegro film than for TK8 film (+75%), and this decrease is therefore not considered as an issue.

Figure 8 Puncture resistance - deflection at maximum load

3.5.1.5 Tear Resistance A slight trend to increase in resistance over time seems apparent for samples in MD, but is not confirmed for samples in TD (especially for lot 1). High variability is classically seen with this test as mentioned before (RSD up to 30%), explaining the differences in size of the confidence intervals.

It is therefore concluded that tear resistance remains equal or slightly increase over time.

Figure 9 Tear resistance - average force

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3.5.1.6 Flex Crack Resistance No significant change is observed during the 25 months aging period. Results remain similar to T0 for all time points, with a clear discrimination between results at 500 and 1000 cycles and results at 5000 cycles.

3.5.2 Summary Shelf-life study on Allegro film shows no significant change over time, except for deflection at rupture. While the load at rupture remains unchanged, the deflection at which rupture occurs decreases by about 25-30% over the 2 years shelf life. This indicates a little less flexible ("ductile") behavior of the film, without significant change in resistance. Such behavior is similarly observed with TK8 film, deflection at rupture decreasing over time. Allegro film deflection at rupture is significantly higher than the TK8 film, presenting a higher resilience to local deformations. Such deformations should not be encountered in normal applications, considering that bag chambers must be kept in their holder, adequately protected from external aggression.

This is confirmed by more than 10 years of experience, as no issues related to a decrease in deflection at puncture have been reported, either for Allegro or TK8 chambers.

3.6 Barrier Properties 3.6.1 Test Data Gas barrier properties of Allegro and TK8 films, obtained on samples gamma-irradiated at 50 kGy, are included in the Table 20. Data for TK8 film have been generated at the time of qualification of the film. Similar data had been generated for Allegro film, but with other testing laboratories and different testing conditions for O2 transmission rate.

To enable optimum comparison, O2 (90% RH inside - 50% RH outside), CO2 (0% RH) and water vapor transmission rates have been retested on Allegro film by the test laboratory used for the TK8 film.

Table 20 Gas barrier properties

Gas Barrier Properties Allegro Film TK8 Film

O2 transmission rate

ASTM D3985 (23°C, 0% RH) <0.05 cm3/(m2.day.bar) Not tested

ASTM F-1927-14 (23 °C, 90% RH inside, 50% RH outside) 0.54 cm3/(m2.day.bar)* 0.52 cm3/(m2.day.bar)

CO2 transmission rate

ASTM F2476-05 (23 °C, 0% RH) < 1 cm3/(m2.day.bar)* < 1 cm3/(m2.day.bar)

Water vapor transmission rate (WVTR)

ASTM F1249-06 (23 °C, 100% RH inside, 0% RH outside) 0.38 g/(m2.day)* 0.40 g/(m2.day)

* Data generated in 2018, by the test laboratory used for TK8 film All data in this table corresponds to steady-state values

O2 (90% RH inside - 50% RH outside) transmission rate measured on 1 sample, CO2 (0% RH) transmission rate measured on 2 samples and water vapor transmission rate measured on 2 samples. In the last 2 cases, the exact same value was obtained.


3.6.2 Comparison Between Films The significant difference between O2 transmission rates obtained with ASTM D3985 and with ASTM F1927 can be attributed to the detrimental effect of moisture on EVOH's gas barrier properties (moisture is reaching EVOH layer by permeating through the LDPE layers). The ASTM D3985 value gives an estimate of the O2 barrier properties for relatively short contact time while the ASTM F-1927 gives one for storage of aqueous-based solutions. The steady-state value for ASTM F-1927 was reached after more than 1500 hours (more than 2 months).

Under ASTM F-1927 test conditions, the O2 transmission rate is comparable between the Allegro and TK8 films, despite the EVOH layer of the Allegro film being twice as thick as that of the TK8 film. The different EVOH material grade found in the TK8 film is less impacted by moisture than the EVOH material in the Allegro film.

3.6.3 Summary Dataset currently available demonstrates that Allegro film barrier properties are equivalent to TK8 film barrier properties. No risk is therefore identified when transitioning from SU systems manufactured with TK8 film to SU systems manufactured with Allegro film.

3.7 Compliance Standards on Raw Materials and Film The data below have been generated with gamma-irradiated samples at the time of initial qualification of the films and are still valid. The BSE/TSE certification is based on certification supplied with the film raw materials.

Table 21 Compliance standards on raw material and film

Allegro Film TK8 Film Remark

USP Biological Reactivity • USP<87> 1

• USP<88> Class VI 1 ; 2

Biological Evaluation of Medical Device • ISO10993-Part 4 - 3

• ISO10993-Part 5 4

• ISO10993-Part 6, 10, 11 4

Plastic for Containers

• USP<661>

• JP XIV Section 61 - Part 1 7 - 3

• EP 3.1.5. 4 ; 5

• EP 3.2.2.1. 6 1 TSE/BSE - Animal Derived Components ADCF certified • EU n° 10/2011 • FDA CFR 21, 177.1520, 178.2010 -

1 Tests have been performed on gamma-irradiated film at 50 kGy minimum 2 According to method described in the USP<88> Class VI, Allegro film was tested at 50 °C and TK8 film tested at 70 °C 3 Tests have been performed on gamma-irradiated film at a dose between 26.6 kGy and 32 kGy

4 Tests have been performed on gamma-irradiated film at a dose between 26.6 kGy and 32 kGy for Allegro film and at 50 kGy minimum for TK8 film 5 Complies with current version of EP 3.1.5. “Polyethylene with additives for containers for parenteral preparations and for ophthalmic preparations”. Inner ULDPE layer of both films has been tested according E.P 6.0 Section 3.1.5.; in both cases, it did fail the test "substances soluble in hexane" because of the low crystallinity of the resin and was considered as not applicable. This test "substances soluble in hexane" is no longer part of current version of Section 3.1.5.

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6 Test recently performed on gamma-irradiated film at a dose between 30 and 50 kGy (part of the new datasets generated for the comparison guide). Result is PASS 7 Replaced by Section 7.02 Part 2.1 in current revision (JP XVII)

Both films have passed standard compliance testing and are compliant to be declared Animal Derived Component Free (ADCF).

No risk is identified when transitioning from SU systems manufactured with TK8 film to SU systems manufactured with Allegro film.

3.8 Extractables Studies on TK8 and Allegro Films In addition to extractables studies performed at the time of initial film qualification, Pall has also conducted additional extraction studies on Allegro film aligned to the current state of the art BPOG2 and draft USP <665> protocols. The Allegro film BPOG and draft USP <665> extraction datasets are the current industry consensus for extractables characterization of single-use components. Pall will continue to support these requirements for single-use components in future. These data will be provided to end users for their information and for input into their risk assessment procedures.

3.8.1 Extraction Conditions Extraction conditions for existing datasets, including solvents, duration, temperatures, surface area to volume ratio and gamma irradiation dose are summarized in Table 22. The controlled report reference containing each of the datasets is indicated at the bottom of Table 22, and can be made readily available with an appropriate confidentiality agreement.

The BPOG-aligned Allegro film datasets aim at covering a large range of bioprocess applications. They are, in general, more aggressive and comprehensive than the TK8 film dataset, based on the solvents used as well as higher surface area to volume ratio.

The draft USP <665> dataset covers a narrower pH range using a single, representative 21-day timepoint, but still covers a large number of bioprocess applications and represents conditions associated with considerably fewer extractables to risk assess.

A historic dataset on Allegro film also exists, which contains additional 90-day extractions in 96% ethanol as well as DMSO.

BPOG-aligned datasets cover 70 days contact time at 40 °C. In case of need, the historic dataset on Allegro covers up to 91 days at 40 °C.

The DCM extraction study is aligned to a now-dated exhaustive extraction approach and is likely far too severe to simulate actual use conditions. It was historically used to generate the broadest list possible of the compounds that can eventually migrate from the film. Similarly, extraction with 96% ethanol is extremely aggressive compared to most biologics applications, whereas 50% ethanol is generally sufficiently organic to cover most actual use conditions.

2 BioPhorum operations group (BPOG) - More information can be found on BPOG website. See https://www.biophorum.com/wp-content/uploads/2016/10/17_file.pdf for the BPOG recommended protocol and the "Best practices guide for evaluating leachables risk from polymeric SU systems used in biopharmaceutical manufacturing" in https://www.biophorum.com/category/media/white-papers/ for the E&L risk assessment.

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Table 22 TK8 and Allegro film extraction conditions

Conditions TK8 Film 1

Allegro Film

BPOG USP<665> Other Historical Data 1

Solvents

Water 1 M NaCl HCl (pH < 3) NaOH (pH > 11) 20% Ethanol 0.1% Polysorbate 80 (PS80) 10% Dimethyl sulfoxide (DMSO) Dichloromethane (DCM)

Water 5 M NaCl 0.1 M H3PO4

(pH ~1.6)

0.5 N NaOH (pH ~13.5) 50% Ethanol 1% PS80

KCl / HCl (pH 3) Phosphate Buffer (pH 10) 50% Ethanol

Water 3 M NaCl Phosphate buffer solution (PBS), pH 3 PBS, pH 11 96% Ethanol 1% PS80 10% DMSO

Contact temperature and duration

DCM: (Reflux, 8 h) DMSO: -20 °C (7 d, 1 and 6 mo.) Others: 40 °C (7 d, 1 and 6 mo.)

25 oC (½ h), 40 oC (24 h, 21 and 70 d) 40 oC (21 d)

96% Ethanol: 25 oC (91 d) DMSO: -20 oC (91 d) Others: 40 oC (91 d)

Surface area to volume ratio DCM: 4.2 cm2/mL Others: 0.56 cm2/mL 6 cm2/mL 6 cm2/mL 0.5 cm2/mL

Number of lots tested 1 2 2 1

Gamma irradiation dose min. of 50 kGy 50 ± 5 kGy 50 ± 5 kGy 27 - 32.5 kGy

Reference USTR3008 VAL-AS-003176-ER (01) VAL-AS-006452-ER (00) USTR2564

1 Data generated at the time of initial qualification

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3.8.2 Analytical Methods The analytical methods used for each of the studies indicated above are highlighted in Table 23. Although each of the reports includes assay for volatiles by headspace GC/MS, semi-volatiles by direct injection GC/MS, non-volatiles LC/MS, and elemental impurities by ICP or OES/MS, the BPOG and USP <665> datasets are intended to provide more robust detection capabilities by including sample concentration steps, LC/UV screening, and non-targeted LC/UV/MS screening.

Table 23 Analytical methods

Extractables TK8 Film

Allegro Film

BPOG and USP<665> Other Historical Data

Volatile organic compounds

Headspace GC/MS Headspace GC/MS Headspace GC/MS

Semi-volatile organic compounds

Direct Injection GC/MS

Direct Injection GC/MS with 20-50 x concentration step

Direct Injection and Derivatization (organic acids) GC/MS

Non-volatile organic compounds

LC/MS (APCI +/-) (targeted screening only)

LC/PDA/MS (ES+/-, APCI +/-) (targeted + non-targeted screening)

LC/MS (APCI +/-) (targeted screening only)

Elemental impurities ICP/OES ICP/MS ICP/OES

Others TOC, pH and conductivity TOC, pH and conductivity

IC (Acetate/Formate), TOC, pH and conductivity

APCI – Atmospheric pressure chemical ionization ES – Electrospray GC - Gas chromatography IC - Ion chromatography ICP - Inductively coupled plasma LC - Liquid chromatography MS - Mass spectrometry PDA - Photodiode array OES - Optical emission spectrometry UV - Ultraviolet detector For Direct injection GC/MS, Liquid-liquid extraction with dichloromethane was performed prior to sample injection

3.8.3 ICH Q3D Elemental Impurities Since the initial qualification of TK8 and Allegro film, new guidance (ICH Q3D) has gone into effect for limits on elemental impurities in final drug product.


Table 24 Summary of the elemental impurities tested evaluated in each of the existing datasets, with the BPOG and USP datasets covering all ICH Q3D specified elemental impurities.

Element ICH Q3D Class TK8 Film

Allegro Film

BPOG and USP <665> Other Historical Data

Cd 1 ● ● ●

Pb 1 ● ● ●

As 1 ●

Hg 1 ●

Co 2A ● ● ●

V 2A ● ●

Ni 2A ● ● ●

Tl 2B ●

Au 2B ●

Pd 2B ●

Ir 2B ●

So 2B ●

Rh 2B ●

Ru 2B ●

Se 2B ●

Ag 2B ● ● ●

Pt 2B ● ●

Li 3 ● ● ●

Sb 3 ●

Ba 3 ● ● ●

Mo 3 ●

Cu 3 ● ● ●

Sn 3 ●

Cr 3 ● ● ●

B N/A ●

Na N/A ● ● ●

W N/A ●

Mg N/A ● ● ●

Al N/A ● ● ●

Ca N/A ● ● ●

Ti N/A ● ●

Mn N/A ● ● ●

Fe N/A ● ● ●

Zn N/A ● ● ●

K N/A ● ● ●

Sr N/A ● ●

Hf N/A ●

Si N/A ●

Zr N/A ●

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3.8.4 Summary of ICH Q3C Residual Solvents and ICH Q3D Elemental Impurities With the relatively recent advent of ICH Q3C residual solvent limits and ICH Q3D elemental impurities limits for drug products, awareness and risk assessment of potential levels of these solvents and elemental impurities in materials that enter, or contact, the drug manufacturing process stream are considered part of a Quality by Design (QbD) process. Table 25 shows the low (sub ppm) levels of ICH Q3C Class 2 residual solvents identified as part of the organic extractables studies, as well as the single ICH Q3D elemental impurity detected above the industry standard reporting threshold. The low levels of the solvents and elemental impurity are not expected to present a risk concern.

Table 25 Summary of ICH Q3C residual solvents and ICH Q3D elemental impurities

Extractables TK8 Film

Allegro Film

BPOG USP<665> Other Historical Data

ICH Q3C Residual solvents

Class 31: Acetic acid Acetone Ethanol Methyl acetate Methyl ethyl ketone (< 120 ppb)

Class 2: Hexane Cyclohexane (< 120 ppb)

None detected

Class 2: Hexane, Cyclohexane (< 1 ppm) Class 3: Acetic acid (from IC) (< 1 ppm)

ICH Q3D Elemental impurities

Class 3: Barium, Lithium (< 10 ppb)

None above 20 ppb (BPOG reporting threshold)

Class 3: Molybdenum (< 70 ppb) None detected

1 All ICH Q3C solvents were detected in DCM extraction samples only

3.8.5 Summary Robust extractables datasets are available for the Allegro film, including those following the BPOG and the draft USP <665> protocols. BPOG and draft USP <665> datasets are available for the film and for the HDPE fittings welded on the film.

Pall recommends privileging these datasets for E&L assessments when detailed extractables profiles are needed, as they correspond to the most recent common industry approach. They should cover most of the applications when transitioning from TK8 to Allegro film.

In case other E&L data are needed to perform E&L risk assessments, please contact your Pall representative.

3.9 Chemical Compatibility It is widely accepted that chemical compatibility is dependent on the chemical structure of the material considered, e.g. LDPE. As product contact layers for both films is ULDPE, a sub-class of LDPE, no difference in chemical compatibility is expected between TK8 and Allegro films.

Besides the fact that no chemical compatibility issue has been identified during the extractables studies described in Section 3.8, some specific chemical compatibility studies have been run on TK8 film, according ASTM D543-06.

Table 26 summarizes the samples' exposure conditions: test solvents, concentration, temperature and contact time.


Table 26 Sample exposure conditions for TK8 film

Product (Test Solvents) Concentration Temperature Contact Time

Acetic acid 100 g/L 60 °C 3 hours

NaOH 50% 200 g/L (5 M) 60 °C 3 hours

MgSO4.7H2O 100 g/L 30 °C 7 days

MnSO4.7H2O 100 g/L 30 °C 7 days

KH2PO4 136 g/L (1 M) 30 °C 7 days

Ethanol 55% 550 mL/L 30 °C 7 days

Bleach (NaClO) 15° + NaOH 0.5 M 50/50 (v/v) 50 °C 3 hours

Riboflavine (vit B2) 10 g/L 30 °C 7 days

Kanamycine sulfate 100 g/L 30 °C 7 days

Caseine hydrolysate 30 g/L 30 °C 7 days

TRIS: Tromethanine (HOCH2)3CNO2 100 g/L 30 °C 7 days

Diethanolamine (DEA/Ureum/SLS) 5 g/L + Ureum 8 M + SLS 1% 30 °C 7 days

Guanidine HCl 800 g/L 30 °C 7 days

Yeast extract 50 g/L 30 °C 7 days

Film resistance to the test solvents was evaluated with the following tests/measurement: film thickness, bag chamber weight, bag chamber dimensions, mechanical tests on film, microscope examination, IR spectra and bag chamber leakage test.

TK8 film was stated resistant to the 14 exposure conditions.

If no new material is in contact with the product further to the migration from TK8 film to Allegro film, no change in chemical compatibility is identified.

4 Manufacturing Process Change Qualification

As part of the TK8 film discontinuation process, a risk assessment was performed by Pall Biotech taking into consideration manufacturing environment, processes and qualifications to define the most appropriate replacement strategy. TK8 bag chambers are manufactured in Pall Life Sciences BVBA in Hoegaarden, Belgium while Allegro bag chambers are currently exclusively manufactured in Pall Medistad BV in Medemblik, The Netherlands. In order to guarantee the continuity of supply, the decision was made to build a new manufacturing cleanroom in Hoegaarden, to enable manufacturing capability for Allegro bag chambers.

Risk assessment was applied at 3 levels:

1. Qualification of a new cleanroom (manufacturing environment) 2. Manufacturing processes of TK8 bag chambers and Allegro bag chambers and SU systems 3. SU systems qualification

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4.1 Manufacturing Environment Pall Life Sciences Belgium BVBA in Hoegaarden invested in a new cleanroom to produce Allegro single-use systems. It is a 250 m² area with filter class Hepa14 and top down air flow. In Q1 2019, the empty cleanroom has been classified as ISO 5 at rest. Current data with equipment installed and operators in the cleanroom confirm the cleanroom will be classified as ISO 7 in routine operations. In Q1 2020, after all equipment and materials installation, the new operational qualification will be performed as well as performance qualification to confirm classification as ISO 5 at rest and ISO 7 in operations. Once the manufacturing process fully validated, for Allegro SU chambers and systems, Pall Life Sciences Belgium BVBA will have a manufacturing cleanroom qualified ISO 7 in operation according to ISO 14644-1 under a Quality System certified to ISO 9001:2015. 4.2 Manufacturing Processes As highlighted in previous chapters, Allegro film and TK8 film have a few different characteristics which prevent the use of Allegro film in the current manufacturing process of TK8 bag chambers. Therefore, the manufacturing processes of Allegro bag chambers, implemented in the new cleanroom of Pall Life Sciences Belgium BVBA facility in Hoegaarden, has been developed to align as much as possible with Pall Medistad BV process, in Medemblik, while reaching equivalent claims and performances to TK8 bag chambers.

The process mapping performed, highlighted differences between the two manufacturing processes and their impact on products performances was evaluated. Those differences are shown in red in the Figure 10 describing Allegro bag chambers and SU systems manufacturing process.

The production of TK8 and Allegro SU systems follow the same production principles with welding of fitments, chamber closure, component assembly, leak testing, packaging and gamma irradiation steps. However, some differences are observed between TK8 and Allegro bag chambers production process as listed below and explained in the next paragraphs:

• Format of film

• Leak testing


Figure 10 Manufacturing process of SU systems with Allegro film

1 The Pall Life Sciences Puerto Rico LLC manufacturing site in Fajardo, Puerto Rico is not manufacturing bag chambers (refer to Section 2.4). Therefore, their manufacturing process starts here with the delivery of bag chambers from either Medemblik, the Netherlands or Hoegaarden, Belgium.

Components (all fitments and films) enter the clean room

Allegro film sheets are cut to shape

Fitments are welded to Allegro sheets

Allegro film sheets are welded

Leak test with pressure decay

equipment

Allegro Biocontainer / chamber closure

System assembly with components (i.e. Tubing, connectors, filters, etc.) 1

Visual inspection of system

Packaging of the SU systems

Gamma irradiation of SU systems (3rd party)

Shipment to end user

Systems returned to Pall Mfg Site (batch recording)

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4.2.1 Differences in the Film Format The raw materials used to create the Allegro chamber is a polymer sheet instead of a polymer body used for TK8 system production (see Figure 11). The first consequence of this modification is cutting the film roll with a validated plotting machine to obtain a polymer sheet with the correct dimensions to create the chamber and holes for the fitments welding. Once the polymer sheets are cut, the fitments are added on the polymer sheets with a welding machine like used by the TK8 process.

The second consequence is the way to close the chamber. In fact, with the TK8 body, the bag chamber is welded using a pre-assembled element while with the Allegro film, separate polymer sheets need to be welded together step by step.

Visually there is a difference in the welding width (Allegro welding being narrower than TK8 welding) but impact on the welding quality is negligible for the following reasons:

• All machines used are based on existing technologies used in other Pall production sites

• The welding process is validated to ensure consistent welding between the different film sheets for the bag chamber creation

• All the bag chambers are leak tested by applying a pressure decay method

Figure 11 Comparison between raw materials design used for bag chamber production

4.2.2 Leak Testing Differences The leak testing is performed on Allegro bag chamber after welding and before assembly while TK8 bag chambers are leak tested after the assembly of the tubing. This difference can be explained due to the fact that Allegro bag chambers have been and are produced in Pall Medistad BV with this approach since 2007, without any reported issue. The Allegro bag chambers production in Pall Life Sciences Belgium BVBA will follow the same production flow.


The impact on product quality is expected negligible for the following reason:

• Junction test is performed on all the fitments, components and tubings during the validation process and only validated items are used in routine production.

• All the bag chambers are leak tested by applying a pressure decay method. The testing parameters will be the same in both, Pall Life Sciences Belgium BVBA and Pall Medistad BV, manufacturing sites.

Table 27 New cleanroom equipment validation status

Equipment Validation Status Date of Completion

Fitment machine On-going Completed

Long weld bar On-going Completed

Plotter On-going Completed

Leak tester On-going Completed

Vacuum sealer 1 On-going Scheduled for June 2020 1 Equipment used for packaging of SU systems

4.2.3 Summary Manufacturing processes of Allegro bag chambers show differences when compared with TK8 products due to differences of properties between the films. Nevertheless, these differences do not impact SU products claims and performances which remain the same whatever film used.

4.3 SU systems Qualifications: New Cleanroom As described in Section 2.4, SU systems can be assembled in one of the three qualified Pall Biotech manufacturing sites, while manufacturing process can differ, all SU systems are qualified according to the same specifications and compliance regardless of the manufacturing site location.

The below flowchart, in Figure 12, describes validation of single-use systems and components performed within all Pall Biotech manufacturing sites. Qualification plans have been defined to demonstrate equivalency between SU systems with chambers made of different films considering different cleanroom environment qualifications and in-process controls.

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Figure 12 Qualifications and in-process controls of SU systems (SUS)

4.3.1 SU systems and Components Qualifications All components in Pall's SU systems are either ADCF or complying with EMA/410/01 rev.3.

Qualification of the SU Systems requires passed junction tests for all connections allowing the release of SU system design for manufacturing. This manufacturing check is performed during design development of each individual SU system.

Data generated below on particulate matter, endotoxins, and bioburden depends on the manufacturing environment of the SU manufacturing site.

Tests are performed at regular intervals for particulate matter, bioburden and endotoxins, according to Pall's manufacturing SOPs. As part of initial manufacturing process qualifications, a master system, including all type of components, is manufactured in standard manufacturing conditions and then tested for fluid path cleanliness (endotoxins and particles level). Results are checked for compliance to USP standards. Due to slight difference in manufacturing process & raw materials (film), the particles profile in the fluid path of TK8 SU systems is different from the particles profile in the fluid path of Allegro SU systems.

A different master system is manufactured to establish the bioburden level of the fluid path for each manufacturing cleanroom. It is used to validate the minimum gamma irradiation dose (VDmax dose) necessary to claim sterility at 10-6 sterility assurance level according to ISO11137 standard.

At the date of qualification package writing, these tests are in progress and the full qualification of the cleanroom in Pall Life Sciences Belgium BVBA in Hoegaarden, Belgium is scheduled for September 2020.

ManufacturingQualifications(Cleanliness, sterility claim,

shelf-life, junctiontest)

Components In-Process Controls

(Integrity, Pressure decay

test, seal strength)

SUS In-Process Controls

(Visual inspection)

Component Qualifications

(ADCF or EMA/410/01rev3,

shelf-life)


A shelf life of 2 years after gamma-irradiation is guaranteed by Pall on the Allegro SU systems. Allegro systems shelf life claims will not be extended beyond 2 years, as it is very likely limited by the shelf life of the other components of the systems. As such, a shorter shelf life may be assigned to systems that contain sensors and/or newly developed components. In these cases, the shelf life of the Pall Allegro SU system will be shortened to not exceed the shelf life of the component with the shortest shelf life.

4.3.2 Standards for SU Systems Containing Chambers Table 28 Standards for SU systems

Standards for SU systems Allegro Film TK8 Film

USP<788> - Particulate matter in injections

USP<85> - Bacterial endotoxins test 1

EMA/410/01 rev.3

ISO 11137 - Sterilization of health care products – radiation 2 3

ISO11737 - Sterilization of health care products -- microbiological methods 2 3

Shelf-life 2 years post gamma-irradiation

1 Routine testing on TK8 and Allegro films is done according USP<85>. Note that USP<85> and EP 3.1.5 standards on endotoxins are considered interchangeable by ICH. 2 and corresponding ANSI/AAMI standards

3 Applicable only to SU systems with a sterility claim

4.3.3 In-Process Controls As part of the qualification of the new cleanroom in Pall Life Sciences Belgium BVBA in Hoegaarden, a risk assessment was performed to ensure equivalency of in-process controls whatever manufacturing site and cleanroom for the Allegro bag chambers manufacturing.

Figure 13 In-process controls of components and SU systems

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4.3.4 Summary SU systems with chambers made with Allegro or TK8 films are produced in compliance to the same standards and offer the same shelf life after gamma-irradiation. The manufacturing environment is also qualified according to same criteria of cleanliness and bioburden level. The manufacturing cleanrooms are certified to the same ISO standards.

Despite different manufacturing processes, the SU systems are submitted to the equivalent in-process controls and they are qualified to reach the same features and specifications. The qualifications show that these differences do not impact product performances.

5 Data Supporting Applications: Performances Assessment

Allegro SU systems used as a replacement of TK8 SU systems have been designed to provide equivalent features and specifications. The maximum operating temperature of + 60 °C was used for testing Allegro storage biocontainers up to 500 L. For higher chamber volumes, Pall Biotech do not recommend using single-use chamber at temperature higher than + 40 °C considering operator safety risk and lack of performance for fluid temperature homogeneity.

Table 29 Operating parameters

Operating Parameters Allegro SU Systems TK8 SU Systems

Temperature

≤ 500 L +2 °C to + 60 °C +2 °C to +60 °C > 500 L +2 °C to + 40 °C

Pressure 0 barg pressure (when inflation < 25 mbar)

Freezing conditions Down to -80 °C very limited handling Down to -70 °C very limited handling

Typical pH ranges Min: 1.0; Max: 13.0

5.1 Mixing Applications 5.1.1 Equivalence of Designs Standard mixing chambers with Allegro film (refer to Section 2.3) were designed according to standard TK8 film based mixing chambers dimensions and using the same mixing method (no change on impeller). Therefore, using an Allegro film version instead of the TK8 film version in the same tank will lead to the same mixing performance.

5.1.2 Durability Tests Table 30 summarizes the durability tests performed at maximum speed to qualify the mixing and bioreactor systems made with Allegro film and using the Allegro impeller.

All systems were checked for leaks during and at the end of the durability tests. Additionally, mechanical tests (tensile and peel tests) were performed on the film and welds after the durability testing. All tests resulted in a PASS result and provide useful data on durability of Allegro film submitted to typical stresses from mixing systems with bottom-located impeller.


Table 30 Durability tests performed on Allegro mixers with Allegro impeller

Size

Duration

Speed

Temperature

Irradiation Dose

Number of samples

50 L

2 days1

200 rpm

2 °C 30-50 kGy 10

Ambient 30-50 kGy 10

40 °C 30-50 kGy 10

200 L

6 days2

150 rpm

2 °C 30-50 kGy 1

Ambient

30-50 kGy 45

50 kGy 9

40 °C 30-50 kGy 1

30 days3 150 rpm 40 °C 50 kGy 9

500 L

6 days2

150 rpm

2 °C 30-50 kGy 1

40 °C 30-50 kGy 1

1000L

6 days2

150 rpm

2 °C 30-50 kGy 1

40 °C 30-50 kGy 1 1 Covering claim of 1 day of continuous mixing 2 Covering claim of 3 days of continuous mixing 3 Covering the duration of a typical cell culture process

5.2 Shipping and Transportation 5.2.1 Intra-Site Transport Verification Transport tests, performed on 1 sample of each volume, confirmed that a loaded 100, 200, 500, 1000 and 1500 L Allegro 3D storage biocontainers in the appropriate Allegro plastic and stainless-steel totes could be transported on its Allegro trolley or castors over a distance of 100 meters without any damage to either the storage biocontainer or the tote.

5.2.2 Shipping Lab Simulations Transport laboratory tests have been performed on Allegro storage biocontainer and shipping stainless steel tote, by TUV (test lab and qualified inspection agency) in Germany. This testing has been done on 200 L and 500 L shipping systems, aligned with criteria of packing group III for testing IBCs intended for use in the transport of dangerous goods (also known as ADR - see standards 30, 31 and 32 in Appendix 1. List of Standards).

Allegro storage biocontainers were filled at full nominal level.

Test sequence includes:

• vibration test • bottom lift test • stacking test • leak test • internal pressure test

All tests results were stated as PASS. More detailed information can be obtained upon request.

Shipping and transportation were not tested with TK8 SU Systems.

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5.3 Freezing Applications 5.3.1 Preliminary Study – TK8 Film A preliminary study has been performed to verify that TK8 2D storage biocontainers could withstand freezing down to -70 °C.

A total of 15 storage biocontainers were stored at -70 °C, for a duration of 2 weeks. The freezing process does not make the film and the tubing more brittle. Twisting and folding them in a frozen state does not create damage. The storage biocontainers are soft and can be manipulated safely in normal conditions. However, the biocontainers cannot withstand any drop, even from a small height. It is advised to protect the biocontainers with a shell designed in a stronger material.

After thawing, traces of delamination of the nylon layer were noticed. However, the storage biocontainers regained their mechanical properties and passed successfully drop and leak testing. The biocontainers integrity is maintained after freezing and thawing.

5.3.2 Preliminary Study – Allegro Film An initial study has been performed to verify that Allegro 2D storage biocontainers withstand freezing down to -80 °C whilst filled at their nominal working volume. Four freeze-and-thaw cycles of 7 days have been performed.

Three storage biocontainers of each size (1 L, 5 L, and 10 L), gamma-irradiated at 50.9 kGy, were filled with water up to their nominal volume and then stored for 7 days at a temperature of -80 °C. After 7 days, they were removed from the freezer and left to thaw at room temperature. When fully thawed, the storage biocontainers were inspected for leaks. They were then placed back into the freezer for another 7 days cycle. The freezing and thawing were performed 4 times, giving 28 days of freezing in total. After 4 times freeze and thaw cycles, the thawed storage biocontainers were dropped from heights according to ASTM D4169-01. The storage biocontainers were dropped both horizontally (twice) and vertically (twice) at this height and were inspected for any sign of water leakages.

The same testing was performed on non-irradiated storage biocontainers, to bracket gamma-irradiation doses.

None of the 1 L, 5 L, and 10 L Allegro 2D storage biocontainers tested showed any signs of water leaks prior to and after the drop testing at a height of 381 mm or 330 mm according to ASTM D4169-01.

5.3.3 Conclusions Both films can withstand freezing down to very low temperatures (tested at -80 °C for Allegro film and -70°C for TK8 film) and require very careful handling in frozen conditions.

After thawing, films recover their performance and storage biocontainers can be handled as usual.


6 Conclusions

Allegro film is an excellent alternative to TK8 film. It is a well-established film that has been used for more than 10 years now.

Its physical characteristics are a bit different: Allegro film is more flexible than TK8 film and can deform more before rupture. It can stick a little more to other components (like tubing) or to the tote's surface. It is therefore recommended to take care of set-up and deployment of the Allegro 3D chambers during the first introduction as an alternative to TK8 chambers, to identify eventual differences in manual handling.

Extensive durability testing data for storage and mixing applications demonstrate that Allegro is perfectly suited for these applications. Freezing preliminary tests gave the same results for both films.

The gas barrier properties of Allegro film are equivalent to the TK8 film properties.

Both films (or SU systems manufactured with these films) have been validated and are monitored according the same compliance standards. No compliance risk is therefore identified.

Complete extractables datasets are available for the Allegro film, including those following the BPOG extractables protocol as well as those following the draft USP <665> protocol.

Chemical compatibility with Allegro film is expected to be the same as TK8 film. The fluid contact layer and the gas barrier layer have the same chemical structure.

Despite manufacturing process differences, SU systems show equivalent features, specifications and performances whatever film is used.

Finally, this change offers a unique opportunity to increase standardization of Allegro SU systems. The design of storage and mixing chambers has been standardized to provide more robust products with Allegro film and reproducible manufacturing methods. While the standard chambers can be used to create customized SU systems for specific needs, the range of Allegro standard SU systems has been extended. This will offer improved guarantee of supply, shorter lead times and optimum quality level.

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Appendix 1. List of Standards

ASTM D882-02 Standard Test Method for Tensile Properties of Thin Plastic Sheeting. This test method covers the determination of tensile properties of plastics in the form of thin sheeting, including film, less than 1.0 mm (0.04 in.) thick.

ASTM D1777-96 (2015) Standard Test Method for Thickness of Textile Materials

FTMS 101C-2065.1 Puncture resistance and elongation test (⅛ in. radius probe method). A conical probe is pushed through the film at a constant speed. The pressure force, needed to break the film, indicates the puncture resistance of the film.

ASTM D1709-01 Standard Test Methods for Impact Resistance of Plastic Film by the Free-Falling Dart Method. This test determines the energy that causes plastic film to fail under impact of a free-falling dart. Weight of the dart, falling from a specified height which would result in 50% failure, is reported. Standard technique (staircase method), method B, is used. For all Allegro samples, a hybrid method had to be used: method B drop height (150 cm) was used with method A dart and weights. Indeed, if sample tested according method B (150 cm drop height, dart B) without weights breaks, the standard requires to use method A (66 cm, dart A) with the weights leading to breakage of the sample. Allegro sample breaks with dart B alone but does not break with dart A and all associated weights. The hybrid method is therefore used.

ASTM F392/F392M-11 Standard Test Method for Flex Durability of Flexible Barrier Materials. This test covers the flex crack resistance of materials by the formation of pinholes. Specimen (sealed film tube) are twisted and horizontally folded at a constant rate and at different test cycles: 500, 1000, 5000 or 10000 test cycles. The number of pinholes is measured by filling the tube with H2O.

ASTM D1004-94A Standard Test Method for Tear Resistance (Graves Tear) of Plastic Film and Sheeting. This test method covers the determination of the tear resistance of flexible plastic film and sheeting at very low rates of loading, 51 mm/min and is designed to measure the force to initiate tearing. Test crosshead speed has been modified from 51 mm/min to 100 mm/min.

ASTM F1927 Standard Test Method for Determination of Oxygen Gas Transmission Rate, Permeability and Permeance at Controlled Relative Humidity Through Barrier Materials Using a Coulometric Detector. This test determines the steady-state rate of transmission of O2 gas through material at a given temperature and relative humidity.

ASTM D3985 Standard Test Method for Oxygen Gas Transmission Rate Through Plastic Film and Sheeting Using a Coulometric Sensor. This test is a sister method of F1927-98, used to determine the rate of transmission of O2 gas through material at a given temperature and in dry conditions.

ASTM F2476-05 Standard Test Method for the Determination of Carbon Dioxide Gas Transmission Rate (CO2TR) Through Barrier Materials Using an Infrared Detector. This test determines the steady-state rate of transmission of CO2 gas through material at a given temperature and relative humidity.

ASTM F1249 Standard Test Method for Water Vapor Transmission Rate Through Plastic Film and Sheeting Using a Modulated Infrared Sensor. This test uses an infrared sensor to measure water vapor permeability at 23 °C with 0% RH on the outside and 100% RH on the inside to simulate worst-case use conditions for a fluid storage container.

USP<87> Biological Reactivity Tests, In Vitro

USP<88> Biological Reactivity Tests, In Vivo

ISO 10993-4 Biological evaluation of medical devices - Part 4: Selection of tests for interactions with blood

ISO 10993-5:1999 Biological evaluation of medical devices - Part 5: Tests for in vitro cytotoxicity


ISO 10993-6:1994 Biological evaluation of medical devices - Part 6: Tests for local effects after implantation

ISO 10993-10:2002 Biological evaluation of medical devices - Part 10: Tests for irritation and skin sensitization

ISO 10993-11:2006 Biological evaluation of medical devices - Part 11: Tests for systemic toxicity

USP<661> Containers - Physicochemical Tests - Plastics

JP XIV - SECTION 61 - Plastic Containers for Aqueous Injections - Part 1. Polyethylene or polypropylene containers for aqueous injections. Moved in Section 7.02 - Part 2.1 for current JP version (JP XVII)

EP 3.1.5. Polyethylene with additives for containers for parenteral preparations and for ophthalmic preparations. Tested initially according EP 6.0

EP 3.2.2.1. Plastic containers for aqueous solutions for parenteral infusion. Tested initially according EP 6.0. UPW is placed in a container and then solutions for parenteral infusion extracted. Appearance of solution, acidity or alkalinity, absorbance, reducing substances and transparency are evaluated.

ASTM F1980-07 Standard Guide for Accelerated Aging of Sterile Barrier Systems for Medical Devices

USP<788> Particulate Matter in Injections

USP<85> Bacterial Endotoxins Test

EP2.6.14. Bacterial Endotoxins

EMA/410/01 REV.3 Effective July 2011, Note for guidance on minimising the risk of transmitting animal spongiform encephalopathy agents via human and veterinary medicinal products

ISO 11137 Sterilization of health care products – Radiation ISO 11137-1:2006 - PART 1 Requirements for development, validation and routine control of a sterilization process for medical devices ISO 11137-2:2013 - PART 2 Establishing the sterilization dose ISO 11137-3:2017 - PART 3 Guidance on dosimetric aspects of development, validation and routine control

ISO 11737 Sterilization of health care products -- Microbiological methods ISO 11737-1 – PART 1 Determination of a population of microorganisms on products ISO 11737-2:2009 - PART 2 Tests of sterility performed in the definition, validation and maintenance of a sterilization process

ASTM D543-06 Standard Practices for Evaluating the Resistance of Plastics to Chemical Reagents

German Executive Order on intrastate and border-crossing Road & Rail Transports of Dangerous Goods [Gefahrgutverordnung Straße und Eisenbahn - GGVSE] as published on 17 June 2009 (BGBl. [Federal Gazette] I, p. 1389).

German Executive Order on Maritime Transports of Dangerous Goods [Gefahrgutverordnung See GGVSee], as published on 3 December 2007 (BGBl. I, p.2815), in particular the International Maritime Dangerous Goods Code (IMDG Code) as amended by the resolution MSC.205(81) in form of its official German translation published on 15 December 2006 (VkBl. 2006, p. 844).

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German Air Transport Licensing Regulations [Luftverkehrszulassungsordnung (LuftVZO)], revised version dated 10 July 2008 (BGBl. I, p. 1229).

ASTM D4169-01 Standard Practice for Performance Testing of Shipping Containers and System


Appendix 2. Comparison of Physical Characteristics Results for TK8 and Allegro Film

Table 31 Comparison of physical characteristics results for TK8 and Allegro films

Physical Characteristics Direction Allegro Film TK8 Film

Tensile Strength at Break MD 15 MPa 31 MPa

TD 13 MPa 34 MPa

Elongation at Break MD 526 % 133 %

TD 450 % 102 %

Elastic Modulus (Young Modulus) MD 293 MPa 486 MPa

TD 295 MPa 478 MPa

Thickness 0.325 mm 0.250 mm

Puncture Resistance

+ Maximum Load 103 N 150 N

+ Deflection 20 mm 13 mm

Dart Impact 193 g 1105 g

Flex Crack Resistance

+ 500 cycles MD 0 pinholes 0 pinholes


+ 1000 cycles MD 0 pinholes 0 pinholes


+ 5000 cycles MD 5.5 pinholes 6 pinholes

TD 5.5 pinholes 4.5 pinholes

Tear Resistance MD 8.6 N 15 N

TD 12 N 22 N

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TK8 and Allegro™ Single-Use Systems and Manufacturing