Abstract
Monoclonal antibodies (mAbs) are the fastest-growing segment in the drug market with eight of the top 20 selling drugs being mAbs and combined sales of close to 60 billion US$/year. The development of new therapeutic mAbs requires the purification of a large number of candidate molecules during initial screenings, subsequent affinity maturation campaigns, and finally the engineering of variants to improve half-life, functionality, or biophysical properties of potential lead molecules. A successful strategy to purify this ever-increasing number of mAbs in a timely manner has been the miniaturization and automation of the purification process using automatic liquid handlers (ALHs) such as Tecan’s Evo or PerkinElmer’s Janus platforms. These systems can be equipped with miniaturized columns, which are available in a wide variety of sizes and affinity matrices to cater to the need of the respective application. Various publications have described the setup of ALHs including the respective purification procedure. However, despite being very precise regarding the overall approach, most publications do not focus on the technical optimization and potential pitfalls, which can be crucial to obtain a robust process. To fill this gap, the present publication is aiming to point at some technical difficulties and suggesting potential ways to overcome these problems in order to facilitate the setup of new ALH systems for the purification of antibodies.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Iervolino A, Urquhart L (2017) World preview 2017, Outlook 2022. www.evaluate.com/PharmaWorldPreview2017
Strohl WR (2014) Antibody discovery: sourcing of monoclonal antibody variable domains. Curr Drug Discov Technol 11(1):3–19
Frenzel A, Hust M, Schirrmann T (2013) Expression of recombinant antibodies. Front Immunol 4:217
Haryadi R, Ho S, Kok YJ, Pu HX, Zheng L, Pereira NA, Li B, Bi X, Goh LT, Yang Y, Song Z (2015) Optimization of heavy chain and light chain signal peptides for high level expression of therapeutic antibodies in CHO cells. PLoS One 10(2):e0116878
Dudgeon K, Rouet R, Christ D (2013) Rapid prediction of expression and refolding yields using phage display. Protein Eng Design Select 26(10):671–674
Dudgeon K, Rouet R, Kokmeijer I, Schofield P, Stolp J, Langley D, Stock D, Christ D (2012) General strategy for the generation of human antibody variable domains with increased aggregation resistance. Proc Natl Acad Sci U S A 109(27):10879–10884
Zalevsky J, Chamberlain AK, Horton HM, Karki S, Leung IW, Sproule TJ, Lazar GA, Roopenian DC, Desjarlais JR (2010) Enhanced antibody half-life improves in vivo activity. Nat Biotechnol 28(2):157–159
Jain NK, Barkowski-Clark S, Altman R, Johnson K, Sun F, Zmuda J, Liu CY, Kita A, Schulz R, Neill A, Ballinger R, Patel R, Liu J, Mpanda A, Huta B, Chiou H, Voegtli W, Panavas T (2017) A high density CHO-S transient transfection system: comparison of ExpiCHO and Expi293. Protein Expr Purif 134:38–46
Schmidt PM, Abdo M, Butcher RE, Yap MY, Scotney PD, Ramunno ML, Martin-Roussety G, Owczarek C, Hardy MP, Chen CG, Fabri LJ (2016) A robust robotic high-throughput antibody purification platform. J Chromatogr A 1455:9–19
Treier K, Hansen S, Richter C, Diederich P, Hubbuch J, Lester P (2012) High-throughput methods for miniaturization and automation of monoclonal antibody purification processes. Biotechnol Prog 28(3):723–732
Wiendahl M, Schulze Wierling P, Nielsen J, Fomsgaard Christensen D, Krarup J, Staby A, Hubbuch J (2008) High throughput screening for the design and optimization of chromatographic processes – miniaturization, automation and parallelization of breakthrough and elution studies. Chem Eng Technol 37(6):893–903
Butcher RE, Martin-Roussety G, Bradford RA, Tester A, Owczarek C, Hardy MP, Chen CG, Sansome G, Fabri LJ, Schmidt PM (2019) Optimizing high throughput antibody purification by using continuous chromatography media. Protein Expr Purif 159:75–82
Chhatre S, Bracewell DG, Titchener-Hooker NJ (2009) A microscale approach for predicting the performance of chromatography columns used to recover therapeutic polyclonal antibodies. J Chromatogr A 1216(45):7806–7815
Ritzen U, Rotticci-Mulder J, Stromberg P, Schmidt SR (2007) Endotoxin reduction in monoclonal antibody preparations using arginine. J Chromatogr B Analyt Technol Biomed Life Sci 856(1–2):343–347
Acknowledgments
The author would like to thank R. Butcher and her team for the continuous interest and enthusiasm in the area of protein purification and characterization, especially while setting up and troubleshooting the protocols for the ALH-mAb purification .
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2021 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Schmidt, P.M. (2021). Technical Optimization for the High-Throughput Purification of Antibodies on Automated Liquid Handlers. In: Labrou, N.E. (eds) Protein Downstream Processing. Methods in Molecular Biology, vol 2178. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0775-6_6
Download citation
DOI: https://doi.org/10.1007/978-1-0716-0775-6_6
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-0774-9
Online ISBN: 978-1-0716-0775-6
eBook Packages: Springer Protocols