PRACTICAL EXP. IIPRACTICAL EXP. II

BIBLIOGRAPHIC WORK

STUDENT TIME SPENT IN THE REPORTBenito Rubio, Alberto 8 hFernández Fernández, Carolina 8 h

IntroductionIntroduction

Obtaining virus clearance is essential in the manufacture of protein-based biopharmaceuticals

This is necessary approval a product for release to market

EfficacyEfficacyDuring the purification process, typically

reduction is:

◦- 103 to 105 monoclonal antibodies per mL

◦- 1010 to 1015 retrovirus-like particles per mL

Virus Removal Virus Removal AlternativesAlternatives

Chemical methods (e.g., solvent detergents, low pH)

physical methods (e.g., heat and radiation)FILTRATION SOMETIMES IT IS THE ONLY POSSIBLE!

MEMBRANE DESIGNMEMBRANE DESIGN

Optimal virus filters must maximize:

acceptable filtrate fluxesreject virus particlesmaximize protein passageObjective:

- capacity- throughput- selectivity

MEMBRANE DESIGN MEMBRANE DESIGN membranes are designed to reject

virus particles while allowing the product of interest to pass through the membrane pores

- Very narrow pore-size distribution- Sturdiness depends on size exclusion - Some layers, need support low pH- Originally tangential flow mode - Today using disposable direct flow

filters- Very narrow pore-size distribution

compared to ultrafiltration membranes

OPERATIONOPERATION-Ultrafiltration membranes, on the other

hand operated with the filtration surface (i.e. skin surface) facing the feed inlet

-membranes are cleaned-in-place and reused

-flux decline is dominated by osmotic pressure effects and gel layer formation

-Asymmetric virus filtration membranes, virus particles often are trapped irreversibly within the more open support

-Compaction and permeability effects

EXAMPLEEXAMPLE

For removal of: parvovirus◦Filters must reject virus particles as

small as 20 nm

Two ultrafiltration membranes have NMWCOs of 300 and 10 kDa.

In addition, virus containing feed streams were spiked with BSA (1% (m/v) final concentration)

PATENTED MEMBRANESPATENTED MEMBRANES

- Normal flow filtration experiments with:- Viresolve 180

- Large pore substructure acts as a depth pre-filter- Declined by nearly 50%

- DV20- 10–20% reduction in permeability

- DV50 - These particles could be removed using a small pore size filter

placed directly in-line

- Omega 300- using different flow orientations- Less dramatic increases in flux

VIRUS REMOVAL BY VIRUS REMOVAL BY FILTRATION:FILTRATION:

ANIMAL VIRUS :ANIMAL VIRUS :

Minute canine parvovirus (CPV), risk of contamination is common to potable water

supply

GVS Speedflow® Positive equipped with a positively charged 0.2 μm membrane. compared with :

Mustang Q®, Speedflow® Positive and Mustang Q®

Results employed virus models: Virus interact with negatively charged components of

cell membranes such as GAGs and sialic acid, Able to attach to positively charged membranes

providing new insights into their electrostatic properties.

PATENTPATENT

-Methods for producing immunoglobulins and in particular anti-D immunoglobulin substantially free of virus and product resulting therefrom.

Specifically provided are methods for nanofiltration of the anti-D immunoglobulin in high ionic strength buffer and with excipient such as polysorbate 80. Additional steps include diafiltration to concentrate the anti-D protein and reduce the concentration of excipient present.

PATENTPATENT

3449314 June 1969 Pollack

3916026 October 1975 Stephan

4021540 May 1977 Pollack et al.

4141887 February 1979 Seufert

4590002 May 1986 Zolton et al.

4880913 November 1989 Doleschel et al.

5115101 May 1992 Bloom et al.

5215681 June 1993 Truong et al.

5723123 March 1998 Karges et al.

U.S. Patent Documents

BIBLIOGRAPHYBIBLIOGRAPHY 1 Understanding virus filtration

membrane performance Journal of Membrane Science, Volume 365,

Issues 1-2, 1 December 2010, Pages 160-169S. Ranil Wickramasinghe, Emily D. Stump, David L. Grzenia, Scott M. Husson, John Pellegrino

  http://www.sciencedirect.com/science?_ob=Miam

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BIBLIOGRAPHYBIBLIOGRAPHY Indexed references [1] P.Y. Huang and J. Peterson, Scaleup and virus

clearance studies on virus filtration in monoclonal antibody manufacture, W.K. Wang, Editor, Membrane Separations in Biotechnology, Marcel Dekker, New York (2001).

[2] A. Higuchi, M. Nemoto, H. Koyama, K. Hirano, B.-O. Yoon, M. Hara, M. Yokogi and S.-I. Manabe, Enhanced microfiltration of γ-globulin solution upon treatment of NaCl addition and/or DNase digestion. J. Membrane Sci.,  210  (2001), pp. 369–378.

[3] T. Ireland, H. Lutz, M. Siwak and G. Bolton, Virus filtration of plasma-derived human IgG: a case study using Vireslove NFP. Biopharm International,  17 11 (2004), pp. 33–40.

…

BIBLIOGRAPHYBIBLIOGRAPHY Indexed references [1] P.Y. Huang and J. Peterson, Scaleup and virus

clearance studies on virus filtration in monoclonal antibody manufacture, W.K. Wang, Editor, Membrane Separations in Biotechnology, Marcel Dekker, New York (2001).

[2] A. Higuchi, M. Nemoto, H. Koyama, K. Hirano, B.-O. Yoon, M. Hara, M. Yokogi and S.-I. Manabe, Enhanced microfiltration of γ-globulin solution upon treatment of NaCl addition and/or DNase digestion. J. Membrane Sci.,  210  (2001), pp. 369–378.

[3] T. Ireland, H. Lutz, M. Siwak and G. Bolton, Virus filtration of plasma-derived human IgG: a case study using Vireslove NFP. Biopharm International,  17 11 (2004), pp. 33–40.

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