Process intensification and continuous bioprocessing by Novasep · 2019. 7. 29. · WHITE PAPER Process intensification and continuous bioprocessing by Novasep 3 21 CFR 210.3 extracts:

Process intensification and continuous bioprocessing by Novasep

W H I T E P A P E R pharmaceuticalsbiopharmaceuticals

bio-industries

agrochemicals

functional ingredients

food ingredients

fine chemicals

biopharmaceuticals

W H I T E P A P E R Process intensification and continuous bioprocessing by NovasepW H I T E P A P E RProcess intensification and continuous bioprocessing by Novasep2

Introduction

Like petroleum, steel, automobile and consumer goods several decades

ago, the biopharmaceutical industry is currently considering a wider

implementation of continuous processing as part of a global process

intensification objective. Continuous processing is already well accepted on

the upstream processing side, since perfusion mode culture has been in use

for decades at an industrial scale.

Justifications for switching from batch to continuous chromatography

are numerous. Besides, the rising demand of biologics suggests reduced

processing and labor costs, footprint reduction, flexibility increase, more

stringent requirements for controlled, more consistent and improved quality;

and most importantly, the requirement for higher productivity and therefore

overall manufacturing cost reduction.

Last but not least regulators are pushing for continuous processing.

About Novasep

Novasep is a leading provider of purification technologies.

It has a unique business model based on 2 sets of offerings. “Process Solutions” with Process

engineering and supply of equipment, and “Manufacturing Solutions” with synthesis and

process development and contract manufacturing (CDMO).

Since decades Novasep designs, develops and operates continuous manufacturing processes.

This is perfectly illustrated by its production facility in Mourenx, France, where is produced

ultra-pure API omega-3 EPA: this is the largest FDA-inspected continuous HPLC Chromatography

Plant in Pharma, worldwide.

Thomas Flouquet, MSc, Application Specialist & Product Manager at Novasep

W H I T E P A P E R Process intensification and continuous bioprocessing by Novasep 3

21 CFR 210.3 extracts:

Batch: a specific quantity of a drug or other material that is intended to have uniform character

and quality, within specified limits, and is produced according to a single manufacturing

order during the same cycle of manufacture.

Lot: a batch, or a specific identified portion of a batch, having uniform character and quality

within specified limits; or, in the case of a drug product produced by continuous process,

it is a specific identified amount produced in a unit of time or quantity in a manner that

assures its having uniform character and quality within specified limits.

Therefore definitions for both “batch” and “lot” are applicable to continuous processes.

FDA views “Though making the switch from batch to continuous manufacturing may be difficult, costly and time consuming, pharma manufacturers and CMOs should begin to consider the switch as in the long-run it will end up saving companies time, money and space”.

Dr. Janet Woodcock, Director of the FDA’s Center for Drug Evaluation and Research. US Congress Hearing,

May 2015.

“Continuous manufacturing is consistent with FDA’s Quality by Design (QBD) efforts”.

FDA Perspective on Continuous Manufacturing. IFPAC Annual Meeting, Baltimore, January 2012.

Sharmista Chatterjee, Ph.D. CMC Lead for QbD ONDQA/CDER/FDA.

W H I T E P A P E R Process intensification and continuous bioprocessing by NovasepW H I T E P A P E RProcess intensification and continuous bioprocessing by Novasep4

Novasep developed the BioSC™ technology for industrial-scale purification of biologics.

BioSC™ is the first technology enabling different multi-columns processes as well as the integration of DSP steps. It allows:

Batch MultiColumn Chromatography (B-MCC)

Sequential MultiColumn Chromatography (S-MCC)

Isocratic MultiColumn Chromatography (I-MCC)

MultiStep MultiColumn Chromatography (MS-MCC)

In addition, single batch chromatography processes can be performed with BioSC™.

BioSC™: Bio System for Chromatography

BioSC™ Pilotexample of 2 MultiColumns set including pump module (left) and columns modules 1 & 2


BioSC™: Bio System for Chromatography B-MCC is the easiest multi-column process since it does not require any

adjustment of the batch recipe. It represents the simplest way to achieve

continuous loading (and if required, elution), by using several columns

operated in parallel that are never connected together.

Batch MultiColumn Chromatography (B-MCC)

Batch chromatography can be represented as a time sequence (chronogram),

presented in Figure 1.

In the following Figure 2 the chronogram of the batch process above will be

applied to a 4-column B-MCC process allowing continuous loading.

B-MCC aims at offering more flexibility. Indeed, BioSC™ Pilot can be configured for one batch module equipped with

one 5 cm ID column or for up to six modules equipped with six 20 cm ID columns. This represents a scale-up factor of

96.

A single system can bring an efficient answer to scale-up or multiproduct plants scenarios.

Consequently, B-MCC simplifies the process design by limiting the number of different systems and columns,

supporting standardization, reducing complexity and offering flexibility.

Another benefit is that B-MCC enables continuous loading and/or elution without any additional process development

nor validation. B-MCC can be easily integrated in any continuous process, encouraging streamlined manufacturing

(Table 1).

Figure 2

Representation of a complete B-MCC cycle (4-column recipe).

As soon as loading is over on column 1,

loading is performed on column 2

As soon as loading is over on column 2, loading is performed

on column 3


on column 4


on column 1

Table 1

Comparison between a standard batch chromatography skid and BioSC™ Pilot configurable from 1 to 6 columns

Figure 1

Example of time sequence for batch chromatography.

Load

Wash 1

Wash 2

Wash 3

Elution

Regeneration

Equilibration

Smallest column possible

Largest column possible

Scale-up factor

Possibility for continuous

loading/ elution

Standard batch chromatography skid on the market (4-180L/h)

5 cm ID 30 cm ID X36 No

BioSC™ Pilot from 1 to 6 column modules (5-100L/h)

5 cm ID 20 cm ID X96 Yes

Performance improvement

IN CONCLUSION: shorter process (streamlined),

higher flexibility and smaller equipment fleet.

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W H I T E P A P E RProcess intensification and continuous bioprocessing by Novasep6

S-MCC process principles:In S-MCC the molecule of interest that breaks through the column is directly loaded onto

the next column that is connected in series. Therefore, loading can be significantly

increased on the first column without losing product.

As soon as the column reaches target saturation, starts the consecutive processing step

i.e. washing, then elution, regeneration, etc... while loading on the next column(s) is ongoing.

This process configuration can be defined as Period 1 (Figure 3).

After equilibration the column can be again loaded and enters Period 2 of the processing

sequence (Figure 4).

Figure 3: Period 1

Elution is performed on a saturated column. Loading is occurring on column 2, 3 and 4.

Figure 4: Period 2

Elution is performed on a saturated column. Loading is occurring on column 3, 4 and 1.

S-MCC is the most well-known example of multi-column process

based on “bind & elute” separation in biopharma. It is a very universal

purification process applicable to all chromatography media, process

solutions and molecules used in batch chromatography. The principle

of S-MCC came from a simple observation: batch chromatography for

both the volume loaded and the loading velocity.

Sequential MultiColumn Chromatography (S-MCC)

Elution Regeneration Equilibration Load

Elution Regeneration Equilibration Load


As soon as all columns of the S-MCC process have gone through the complete processing

sequence, one cycle is completed; a chronogram visualizes utilization of columns

throughout the processing time (Figure 5). S-MCC is based on countercurrent processing

flow of the stationary and liquid phases, and therefore allows for much more efficient

processing and for homogeneous exposure of the stationary phase to the loads compared

to batch, where most of the molecules are mainly exposed to the top section of the packed

column during loading, resulting in concentration gradients.

Figure 5

“Chronogram” (from BioSC™ Predict simulation software) of a complete S-MCC cycle (4-column recipe).

Process dimensioningFigures 6 and 7 help to quickly dimension the S-MCC process of an antibody

capture step based on the feed volume, titer and the processing time requirements.

They illustrate typical processing scenarios for a 4-column recipe depending on

the column ID.

1

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10 g/L

5 g/L

2 g/L

1 g/L

0,5 g/L

10 g/L5 g/L

2 g/L1 g/L0,5 g/L

10 g/L

5 g/L

2 g/L

1 g/L

0,5 g/L

200 L100 L

50 L

Figure 6

2

CON

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OVA

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Figure 7

10 g/L

5 g/L

2 g/L

1 g/L

0,5 g/L

10 g/L5 g/L

2 g/L1 g/L0,5 g/L

10 g/L

5 g/L

2 g/L

1 g/L

0,5 g/L

2000 L1000 L

500 L

W H I T E P A P E RProcess intensification and continuous bioprocessing by Novasep8

Figure 6

Processing time for a BioSC™ 4-column (5cm ID) recipe, depending on feed titer and volume to be processed.

For example, 4 columns of 5cm ID can process 100L of 2g/L harvest in 15h.

Or 4 columns of 5cm ID can process 200L of 10g/L harvest in 92h.

Figure 7

Processing time for a BioSC™ 4-column (20cm ID) recipe, depending on feed titer and volume to be processed.

For example, 4 columns of 20cm ID can process 1000L of 5g/L harvest in 17h max.

Or 4 columns of 20cm ID can process 2000L of 10g/L harvest in 57h max.

Volume of the bioreactor

Volume of the bioreactor

Feed titer

Feed titer


After the definition of the CQAs, different recipes (combination of process parameters)

pre-defined with the BioSC™ Predict process simulation software were tested and performance

results were compared to their reference batch process. An optimized S-MCC recipe was

performed on a 4-column BioSC™ Lab system with a load of a monoclonal antibody solution at

a concentration of 4.5 mg/mL. Same buffers and loads were applied for batch and S-MCC runs.

The following table 2 summarizes the main results of the study. BioSC™ run in S-MCC mode

exhibits important performance improvements compared to the batch process.

Product recovery is similar between Batch and S-MCC recipe.

Aggregates level is comparable.

Higher purity is reached with S-MCC recipe.

HCP (host cell protein) reduction rate is improved with S-MCC recipe.

Less protein A leaching was measured for S-MCC recipe.

DNA reduction rate is in the same range.

Productivity (expressed as mg of MAb/mL of resin/hour) gain is +200% for S-MCC.

Product loss in flowthrough is considered similar.

Columns positioned in series enable one to be loaded at a higher feed amount in a shorter time,

boosting performance of a single chromatography step. As an average, productivity increases by

200% (so X3), cost of goods (buffers and resins) are reduced by 3 to 4 times while CQAs remain in

the same range.

Critical Quality Attributes (CQAs) and process performance study

Sequential multicolumn chromaography (S-MCC)

Table 2

Results of a CQA and process performance study. S-MCC recipe set on BioSC™ Lab and comparison with batch.

Performance in the order of batch results (no color)

IN SUMMARY: shorter processing time, higher productivity

and smaller costs & volumes.

Recovery

(%) Aggregates

(%)Purity

(%)Reduction rate HCP

Protein A leaching (ppm)

DNA Log reduction

Productivity (mg/mL/h)

Loss flowthrough

(%)

Batch-MAb 4.5mg/mL

88 2.6 94 X47 1.6 2 14.6 0.3

BioSC-MAb 4.5mg/mL

89 2.9 98 X147 0 2 44.8 0.25


W H I T E P A P E RProcess intensification and continuous bioprocessing by Novasep1 0

I-MCC aims at rehabilitating SEC at industrial scale. In I-MCC processes, as in S-MCC ones,

several short columns (e.g.: with 10cm bed height) are operated in a countercurrent mode

and therefore increase significantly process performance together with buffer reduction.

The following Figure 8 represents the concentration profile of two different populations

(blue and red) in a batch column during the separation (t1 and t2) and during elution (t3).

Red population includes species with a higher molecular weight than blue population and

therefore elutes earlier.

Size exclusion chromatography (SEC) is known to be a powerful separation technique

for analytical chromatography. But implementing SEC at industrial scale is a challenge.

It suffers from a lack of productivity and high buffer consumption. Besides this, it requires

long beds difficult to pack and manage at large scale.


Figure 8

Batch SEC chromatography separation principle.

t1 t2 t3Pure red population elution

W H I T E P A P E R Process intensification and continuous bioprocessing by Novasep 1 1

In the following Figures 9 to 12 the sequences of a typical 2 populations separation

applying the I-MCC process are illustrated.

Figure 9: I-MCC Sequence 1

By positioning columns in series, no need to wait for complete separation.

Red collection starts as soon as red population elutes from column1.


Mixed zone (containing red and blue populations) is redirected on column2.

Fresh feed is loaded on column2 at the same time.


Pure blue population eluting from column1. Separation occurring on column2.


Period 2: Pure red population collection from column2.

On next sequence, mixed zone (containing red and blue populations) will be redirected on column1.

Fresh feed will be loaded on column 1. Then blue population eluting from column2.

Fresh feed


In the following figure a chronogram represents utilization of columns throughout

the processing time of an I-MCC process (Figure 13).

Column 1 in series with column 2 Blue population collection from column 2

Column 2 in series with column 1 (Figure 11) Blue population collection from column 1

Column 1 alone (Figure 9)Red population collection from column 1

Column 2 alone (Figure 12)Red population collection from column 2

1

2

1

0

2

Fresh feed loading (Figure 10). Column 1 in series with column 2

Fresh feed loading. Column2 in series with column 1

I-MCC recipe was tested to purify a virus. Performance results were compared to their

reference batch process. The tests were performed on a BioSC™ Lab system with 2 columns

of 10 cm bed height. The same SEC media and buffer were used for batch and I-MCC test runs.

The following table 3 summarizes the main results of the study. BioSC™ run in I-MCC mode

exhibits much better performance than batch.

As for S-MCC, I-MCC enables one to load a higher amount of feed in less time, boosting the

performance of a SEC separation. As an average, productivity increases approximately by

5 to 6-fold while reducing buffer consumption by 85%. And last but not least, column size

can be reduced significantly, i.e. -96% SEC media. Only 2X10 cm (bed height) columns are

required compared to 1Xca.80 cm (bed height) column for a batch process.

Figure 13

Chronogram of a complete I-MCC cycle (2-column recipe).

Table 3

Results of process performance study. An optimized I-MCC recipe set on BioSC™ Lab and comparison with batch.

IN CONCLUSION: shorter processing time, higher productivity

and smaller costs & volumes.


Productivity improvement factor

Buffer consumption (20L virus solution

purified in 17h)

SEC media (20L virus solution

purified in 17h)

Batch Virus SEC 1 1700 L 200 L

I-MCC Virus SEC X5.5 256 L 8 L



With one 3-module BioSC™ equipment, 3 chromatography

separations including a viral inactivation (VI) and in-line

dilution (ILD) can be operated simultaneously and continuously.

Straightforwardly, to switch from several batch skids to a single

DSP skid. Such a process has been made possible with BioSC™

technology in a cGMP environment (Picture 1).

Until now, one chromatography system was needed to execute one

chromatography step. Usually, standard processes involve three

chromatography columns, thus three systems.

MultiStep MultiColumn Chromatography (MS-MCC)

Picture 1

BioSC™ Pilot running in MS-MCC mode for the purification of a monoclonal antibody.


The following illustration in Figure 14 shows how MS-MCC integrates chromatography steps

and how the upstream load can be treated in several cycles.

One single skid being able to perform the whole downstream process, from capture to

polishing, represents a drastic footprint reduction, and a serious cost reduction. One of

the key benefits is the elimination of non-added value steps (e.g. intermediate storage),

human intervention (safety, batch failure…) and equipment such as holding tanks.

Data for a 3-chromatography step process, comparing batch to MS-MCC process including

viral inactivation with BioSC™ are available (Table 4).

Productivity is increased by ca. X4 with MS-MCC.

Process time is reduced by 66% with MS-MCC.

Media consumption is reduced by 70% with MS-MCC.

Buffer consumption is reduced by 30% with MS-MCC.

Footprint is reduced by more than 50% with MS-MCC.

A closed DSP prevents human error and potential batch failure.

Figure 14

Representation of MS-MCC run, integrating 4 steps (3 chromatography steps + viral inactivation) of a MAb DSP.

ProtA = Protein A media; VI = viral inactivation, AEX = anion exchanger media


Table 4

Comparison between a standard batch process and a BioSC™ MS-MCC process.

Productivity (mg/mL/h)

Time reduction

(days)

Media consumption

(L)

Buffer consumption

(L)

Footprint reduction

Human error prevention

Standard batch process

2.7 3 20 100 - -

MS-MCC 10.5 1 6 70 >50% +++

All in all, shorter process (streamlined), higher productivity

and safety, and smaller costs and footprint.


VI

Pro

A

AE

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Mix

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VI

Pro

A

AE

X

Mix

edm

ode

VI

Pro

A

AE

X

Mix

edm

ode

VI

VI

Cycle 1

Cycle 1

Cycle3 starts as soon as ProtA/Cycle2

is eluted


is eluted

All 4 steps are run in parallel


is eluted

Cycle 1

Cycle 1

Cycle 1

Time


BioSC™ Predict:

Models thermodynamics and kinetics of your chromatography batch process.

Suggests different MultiColumn Chromatography scenarios depending on your objectives.

Analyses performances of your optimized scenario.

Programs your BioSC™ unit.

Enables scale-up “prediction”.

Deeper process understanding is becoming more and more important to design a chromatographic separation that consistently delivers high quality products. With the process characterization method combined with the BioSC™ Predict software this can be achieved within a minimum of time. Furthermore, whatever BioSC™ technology is applied, process simulation is key for a successful, quick and simple implementation. Novasep designed its own simulation software, BioSC™ Predict, based on algorithms and real-case industrial data.

BioSC™ Predict Software

W H I T E P A P E R Process intensification and continuous bioprocessing by NovasepW H I T E P A P E RProcess intensification and continuous bioprocessing by Novasep1 6

Conclusion

The rising demand of biologics combined with the need of deeper process

understanding, reduced development times and an ever-increasing

pressure on costs compels the Biotech industry to transform its production

models. Modernization of manufacturing tools includes the switch towards

continuous/intensified downstream processes assuring a maximum

of control by applying process and analytical tools.

BioSC™ Lab

BioSC™ Pilot

BioSC™ Predict


BioSC™ technology (B-MCC, S-MCC, I-MCC, MS-MCC and Predict software)

coupled with increased process understanding perfectly match all these objectives.

“The science exists to enable continuous manufacturing of pharmaceuticals. There are no regulatory hurdles for implementing continuous manufacturing. FDA supports the implementation of continuous manufacturing using a science and risk-based approach.”

Sharmista Chatterjee, Ph.D. IFPAC Annual Meeting (Baltimore, January 2012).

The objective is threefold:

> Shorter, streamlined processes.

> Higher productivity, flexibility and quality (through process understanding and control).

> Smaller costs, volumes and footprint.

EUROPE Novasep ProcessSite Eiffel 81 boulevard de la Moselle - BP 50 54340 Pompey - FRANCEPhone: +33 383 49 70 00 Fax: +33 383 49 70 02

NORTH AMERICA Novasep, LLC23 Creek Circle Boothwyn, PA 19061 - USAPhone: +1 610 494 0447 ext. 30 Fax: +1 610 494 1988

ASIA Novasep AsiaBuilding No.4, Lane 1690, Zhangheng Road, Zhangjiang Hi-Tech Park, Shanghai 201203 - CHINAPhone: +86 (0)21 6045 1600 Fax: +86 (0)21 6045 1699

INDIA Novasep IndiaFour Square Elegant, Block A, Flat 110 Madhavapuri Hills, Road5, Hyderabad - 500050 - INDIA Phone: +91 9866573377

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© 2018 Groupe Novasep SAS. Groupe Novasep SAS and/or its affiliates (hereafter Groupe Novasep) own or license the copyrights, trademarks, names, logos, and other rights to the information in this brochure. No right or license is granted to any other losses for access or reliance and not from access. Any unauthorized use without the express prior written consent of Groupe Novasep is prohibited. Disclaimers: The information contained in this brochure are provided “as is”, for informational purposes only, without any representation or warranty of accuracy or completeness. In no event will Groupe Novasep be liable to any party for any damages or any other losses from access or reliance upon any information contained in this brochure.

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Process intensification and continuous bioprocessing by Novasep · 2019. 7. 29. · WHITE PAPER Process intensification and continuous bioprocessing by Novasep 3 21 CFR 210.3 extracts: