Abstract
Protein aggregation is a common issue encountered during manufacture of biotherapeutics. It is possible to influence the amount of aggregate produced during the cell culture and purification process by carefully controlling the environment (eg, media components) and implementing appro-priate strategies to minimize the extent of aggregation. Steps to remove aggregates have been successfully used at a manufacturing scale. Care should be taken when developing a process to monitor the compatibility of the equipment and process with the protein to ensure that potential aggregation is minimized.
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References
US Pharmacopeia. USP/NF General Chapter<788> Particulate Matter in Injections. In: U.S. Pharmacopeia, ed.National Formulary, USP29-NF-24 (suppl 2). Rockville, MD: USP; 2006.
Andya JD, Hsu CC, Shire SJ. Mechanisms of aggregate formation and carbohydrate excipient stabilization of lyophilized humanized monoclonal antibody formulations.AAPS PharmSci. 2003;5:E10.
Shahrokh Z, Sluzky V, Stratton PR, Eberlein GA, Wang YJ. Disulfide-linked oligomerization of basic fibroblast growth factors: effect of sulfated compounds. In:Formulation and Delivery of Proteins and Peptides, ACS Symposium Series 567. Washington, DC: American Chemical Society, 1994:85–99.
Creed D. The photophysics and photochemistry of the near-UV absorbing amino acids. II. Tyrosine and its simple derivatives.Photochem Photobiol. 1984;39:563–575.
Giulivi C, Davies KJA. Dityrosine: a marker for oxidatively modified proteins and selective proteolysis. In: Packer L, ed.Methods in Enzymology. vol. 233. New York, NY: Academic Press; 1994:363–371.
Moore JM, Patapoff TW, Cromwell MEM. Kinetics and thermodynamics of dimer formation and dissociation for a recombinant humanized monoclonal antibody to vascular endothelial growth factor.Biochemistry. 1999;38:13960–13967.
Liu J, Nguyen MD, Andya JD, Shire SJ. Reversible self-association increases the viscosity of a concentrated monoclonal antibody in aqueous solution.J Pharm Sci. 2005;94:1928–1940.
Maislos M, Bialer M, Mead PM, Robbins DC. Pharmacokinetic model of circulating covalent aggregates of insulin.Diabetes. 1988;37:1059–1063.
Rosenberg AS. Effects of protein aggregates: an immunologie perspective.AAPS J. 2006;8:E501-E507.
Fernández A. What factor drives the fibrillogenic association of β-sheets?FEBS Lett. 2005;579:6635–6640.
Chi EY, Krishnan S, Randolph TW, Carpenter JF. Physical stability of proteins in aqueous solution: mechanism and driving forces in nonnative protein aggregation.Pharm Res. 2003;20:1325–1336.
Zhang YB, Howitt J, McCorkle S, Lawrence P, Springer K, Freimuth P. Protein aggregation during overexpression limited by peptide extensions with large net negative change.Protein Expr Purif. 2004;36:207–216.
Freimuth P, Springer K, Berard C, Hainfeld J, Bewley M, Flanagan J. Coxsackievirus and adenovirus receptor amino-terminal immunoglobulin V-related domain binds adenovirus type 2 and fiber knob from adenovirus type 12.J Virol. 1999;73:1392–1398.
Frand AR, Cuozzo JW, Kaiser CA. Pathways for protein disulphide bond formation.Trends Cell Biol. 2000;10:203–210.
Zhang W, Czupryn MJ. Free sulfhydryl in recombinant monoclonal antibodies.Biotechnol Prog. 2002;18:509–513.
Chaderjian WB, Chin ET, Harris RJ, Etcheverry TM. Effect of copper sulfate on performance of a serum-free CHO cell culture process and the level of free thiol in the recombinant antibody expressed.Biotechnol Prog. 2005;21:550–553.
Phillips J, Drumm A, Harrison P, et al. Manufacture and quality control of CAMPATH-1 antibodies for clinical trials.Cytotherapy. 2001;3:233–242.
Ejima D, Yumioka R, Tsumoto K, Arakawa T. Effective elution of antibodies by arginine and arginine derivatives in affinity column chromatography.Anal Biochem. 2005;345:250–257.
Ansaldi D, Lester P, inventors. Genentech, Inc., assignee. Separation of polypeptide monomers. US patent 6 620 918. September 16, 2003.
van Reis R, Zydney A. Membrane separations in biotechnology.Curr Opin Biotechnol. 2001;12:208–211.
Harris RJ, Shire SJ, Winter C. Commercial manufacturing scale formulation and analytical characterization of therapeutic recombinant antibodies.Drug Dev Res. 2004;61:137–154.
van Reis R, Goodrich EM, Yson CL, Frautschy LN, Dzengeleski S, Lutz H. Linear scale ultrafiltration.Biotechnol Bioeng. 1997;55:737–746.
Wan Y, Vasan S, Ghosh R, Hale G, Cui Z. Separation of monoclonal antibody alemtuzumab monomer and dimers using ultrafiltration.Biotechnol Bioeng. 2005;90:422–432.
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Published: September 15, 2006
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Cromwell, M.E.M., Hilario, E. & Jacobson, F. Protein aggregation and bioprocessing. AAPS J 8, 66 (2006). https://doi.org/10.1208/aapsj080366
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DOI: https://doi.org/10.1208/aapsj080366