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Insight into the Purification Strategies for Removing the Byproducts of Bispecific Antibodies |
Qian LI,Xiao-ying LIANG,Guo-zhu LI,Qing-quan HE,Huang-hong TAN,Zi-chen WANG,Guang-liu XU,jing LI,Meng-ni FAN,Dan XU**() |
Nanjing China Tai Tianqing Pharmaceutical Co., Ltd, Fanghua Pharmaceutical Research Institute, Department of Biology, Nanjing 210046, China |
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Abstract Bispecific antibodies can simultaneously bind to two targets. Compared with monospecific antibodies, they have the advantages of high efficacy and less toxic and side effects, so they have become a research hotspot in recent years.However, since bispecific antibodies are composed of different heavy chains and light chains, and the expression of heavy and light chains is difficult to control at the same level, it is very easy to form by-products, which greatly increase the difficulty and cost of downstream purification.In recent years, several pharmaceutical companies have developed bispecific antibody preparation platforms, which have greatly improved the success rate of bispecific antibody assembly.However, various double-antibody molecular design strategies are not enough to completely prevent the production of by-products, so various chromatographic methods are needed to further remove the by-products of double-antibody molecules to improve product quality.This paper reviews several mainstream bispecific antibody design platforms in recent years, and systematically summarizes the chromatographic methods for removing homodimers, half antibodies and 3/4 antibodies and aggregates. The comprehensive information should be helpful to provide a theoretical basis for bispecific antibody purification.
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Received: 23 June 2022
Published: 04 November 2022
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Corresponding Authors:
Dan XU
E-mail: 1563225340@qq.com
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Cite this article:
Qian LI,Xiao-ying LIANG,Guo-zhu LI,Qing-quan HE,Huang-hong TAN,Zi-chen WANG,Guang-liu XU,jing LI,Meng-ni FAN,Dan XU. Insight into the Purification Strategies for Removing the Byproducts of Bispecific Antibodies. China Biotechnology, 2022, 42(10): 60-69.
URL:
https://manu60.magtech.com.cn/biotech/10.13523/j.cb.2206043 OR https://manu60.magtech.com.cn/biotech/Y2022/V42/I10/60
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|
[1] |
Lim S M, Pyo K H, Soo R A, et al. The promise of bispecific antibodies: clinical applications and challenges. Cancer Treatment Reviews, 2021, 99: 102240.
doi: 10.1016/j.ctrv.2021.102240
|
|
|
[2] |
Moek K L, de Groot D J A, de Vries E G E, et al. The antibody-drug conjugate target landscape across a broad range of tumour types. Annals of Oncology, 2017, 28(12): 3083-3091.
doi: 10.1093/annonc/mdx541
pmid: 29045509
|
|
|
[3] |
Gera N. The evolution of bispecific antibodies. Expert Opinion on Biological Therapy, 2022, 22(8): 945-949.
doi: 10.1080/14712598.2022.2040987
|
|
|
[4] |
Esfandiari A, Cassidy S, Webster R M. Bispecific antibodies in oncology. Nature Reviews Drug Discovery, 2022, 21(6): 411-412.
doi: 10.1038/d41573-022-00040-2
pmid: 35246638
|
|
|
[5] |
Linke R, Klein A, Seimetz D. Catumaxomab. mAbs, 2010, 2(2): 129-136.
doi: 10.4161/mabs.2.2.11221
pmid: 20190561
|
|
|
[6] |
Kantarjian H, Stein A, Gökbuget N, et al. Blinatumomab versus chemotherapy for advanced acute lymphoblastic leukemia. The New England Journal of Medicine, 2017, 376(9): 836-847.
doi: 10.1056/NEJMoa1609783
pmid: 28249141
|
|
|
[7] |
Scott L J, Kim E S. Emicizumab-kxwh: first global approval. Drugs, 2018, 78(2): 269-274.
doi: 10.1007/s40265-018-0861-2
pmid: 29357074
|
|
|
[8] |
Davda J, Declerck P, Hu-Lieskovan S, et al. Immunogenicity of immunomodulatory, antibody-based, oncology therapeutics. Journal for Immunotherapy of Cancer, 2019, 7(1): 105.
doi: 10.1186/s40425-019-0586-0
pmid: 30992085
|
|
|
[9] |
Zarrineh M, Mashhadi I S, Farhadpour M, et al. Mechanism of antibodies purification by protein A. Analytical Biochemistry, 2020, 609: 113909.
doi: 10.1016/j.ab.2020.113909
|
|
|
[10] |
Chen S W, Zhang W. Current trends and challenges in the downstream purification of bispecific antibodies. Antibody Therapeutics, 2021, 4(2): 73-88.
doi: 10.1093/abt/tbab007
pmid: 34056544
|
|
|
[11] |
Li Y F. A brief introduction of IgG-like bispecific antibody purification: methods for removing product-related impurities. Protein Expression and Purification, 2019, 155: 112-119.
doi: S1046-5928(18)30607-7
pmid: 30513344
|
|
|
[12] |
Li Y F. Immunoglobulin-binding protein-based affinity chromatography in bispecific antibody purification: functions beyond product capture. Protein Expression and Purification, 2021, 188: 105976.
doi: 10.1016/j.pep.2021.105976
|
|
|
[13] |
Ridgway J B B, Presta L G, Carter P. ‘Knobs-into-holes’ engineering of antibody CH3 domains for heavy chain heterodimerization. Protein Engineering, Design and Selection, 1996, 9(7): 617-621.
doi: 10.1093/protein/9.7.617
|
|
|
[14] |
Schaefer W, Regula J T, Bähner M, et al. Immunoglobulin domain crossover as a generic approach for the production of bispecific IgG antibodies. Proceedings of the National Academy of Sciences of the United States of America, 2011, 108(27): 11187-11192.
|
|
|
[15] |
Brinkmann U, Kontermann R E. The making of bispecific antibodies. mAbs, 2017, 9(2): 182-212.
doi: 10.1080/19420862.2016.1268307
pmid: 28071970
|
|
|
[16] |
Sampei Z, Igawa T, Soeda T, et al. Identification and multidimensional optimization of an asymmetric bispecific IgG antibody mimicking the function of factor VIII cofactor activity. PLoS One, 2013, 8(2): e57479.
doi: 10.1371/journal.pone.0057479
|
|
|
[17] |
Fischer N, Elson G, Magistrelli G, et al. Exploiting light chains for the scalable generation and platform purification of native human bispecific IgG. Nature Communications, 2015, 6: 6113.
doi: 10.1038/ncomms7113
pmid: 25672245
|
|
|
[18] |
Wranik B J, Christensen E L, Schaefer G, et al. LUZ-Y, a novel platform for the mammalian cell production of full-length IgG-bispecific antibodies. Journal of Biological Chemistry, 2012, 287(52): 43331-43339.
doi: 10.1074/jbc.M112.397869
pmid: 23118228
|
|
|
[20] |
Li Y F. IgG-like bispecific antibody platforms with built-in purification-facilitating elements. Protein Expression and Purification, 2021, 188: 105955.
doi: 10.1016/j.pep.2021.105955
|
|
|
[20] |
Wang C L, Vemulapalli B, Cao M Y, et al. A systematic approach for analysis and characterization of mispairing in bispecific antibodies with asymmetric architecture. mAbs, 2018, 10(8): 1226-1235.
doi: 10.1080/19420862.2018.1511198
pmid: 30153083
|
|
|
[21] |
Shiraiwa H, Narita A, Kamata-Sakurai M, et al. Engineering a bispecific antibody with a common light chain: identification and optimization of an anti-CD3 Epsilon and anti-GPC3 bispecific antibody, ERY974. Methods, 2019, 154: 10-20.
doi: S1046-2023(18)30096-3
pmid: 30326272
|
|
|
[22] |
de Nardis C, Hendriks L J A, Poirier E, et al. A new approach for generating bispecific antibodies based on a common light chain format and the stable architecture of human immunoglobulin G1. Journal of Biological Chemistry, 2017, 292(35): 14706-14717.
doi: 10.1074/jbc.M117.793497
pmid: 28655766
|
|
|
[23] |
Moretti P, Skegro D, Ollier R, et al. BEAT® the bispecific challenge: a novel and efficient platform for the expression of bispecific IgGs. BMC Proceedings, 2013, 7(S6): 9.
|
|
|
[24] |
Skegro D, Stutz C, Ollier R, et al. Immunoglobulin domain interface exchange as a platform technology for the generation of Fc heterodimers and bispecific antibodies. Journal of Biological Chemistry, 2017, 292(23): 9745-9759.
doi: 10.1074/jbc.M117.782433
pmid: 28450393
|
|
|
[25] |
Dietrich S, Gross A W, Becker S, et al. Constant domain-exchanged Fab enables specific light chain pairing in heterodimeric bispecific SEED-antibodies. Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics, 2020, 1868(1): 140250.
doi: 10.1016/j.bbapap.2019.07.003
|
|
|
[26] |
Jang S, Song J, Kim N, et al. Development of an antibody-like T-cell engager based on VH-VL heterodimer formation and its application in cancer therapy. Biomaterials, 2021, 271: 120760.
doi: 10.1016/j.biomaterials.2021.120760
|
|
|
[27] |
Steven M. Chamow P P, Pratap L A. Capture of CH1-containing bispecific antibodies. BioProcess International, 2020, 18(5):130-144.
|
|
|
[28] |
Tustian A D, Endicott C, Adams B, et al. Development of purification processes for fully human bispecific antibodies based upon modification of protein A binding avidity. mAbs, 2016, 8(4): 828-838.
doi: 10.1080/19420862.2016.1160192
pmid: 26963837
|
|
|
[29] |
Qin T T, Wang Y, Li Y F. Separating antibody species containing one and two kappa light chain constant region by KappaSelect affinity chromatography. Protein Expression and Purification, 2020, 171: 105618.
doi: 10.1016/j.pep.2020.105618
|
|
|
[30] |
Chen X J, Wang Y, Wang Y, et al. Protein L chromatography: a useful tool for monitoring/separating homodimers during the purification of IgG-like asymmetric bispecific antibodies. Protein Expression and Purification, 2020, 175: 105711.
doi: 10.1016/j.pep.2020.105711
|
|
|
[31] |
Sharkey B, Pudi S, Wallace Moyer I, et al. Purification of common light chain IgG-like bispecific antibodies using highly linear pH gradients. mAbs, 2017, 9(2): 257-268.
doi: 10.1080/19420862.2016.1267090
pmid: 27937066
|
|
|
[32] |
Zhang L, Parasnavis S, Li Z J, et al. Mechanistic modeling based process development for monoclonal antibody monomer-aggregate separations in multimodal cation exchange chromatography. Journal of Chromatography A, 2019, 1602: 317-325.
doi: S0021-9673(19)30580-1
pmid: 31248584
|
|
|
[33] |
Tang J Q, Zhang X D, Chen T, et al. Removal of half antibody, hole-hole homodimer and aggregates during bispecific antibody purification using MMC ImpRes mixed-mode chromatography. Protein Expression and Purification, 2020, 167: 105529.
doi: 10.1016/j.pep.2019.105529
|
|
|
[34] |
Zhang Y, Cai L L, Wang Y, et al. Processing of high-salt-containing protein A eluate using mixed-mode chromatography in purifying an aggregation-prone antibody. Protein Expression and Purification, 2019, 164: 105458.
doi: 10.1016/j.pep.2019.105458
|
|
|
[35] |
Nicolas F, Franois D J, Keith W, et al. methods of purifying bispecific antibodies:EPO,EP3268390A1.2018-01-17[2018-01-17]. https://wenku.baidu.com/view/47f290b8de3383c4bb4cf7ec4afe04a1b071b0f6?fr=xueshu.
|
|
|
[36] |
Qian G. Purification of monoclonal antibodies:USA,WO2011005818A1.2011-01-13[2011-01-13]. https://wenku.baidu.com/view/06a03656f142336c1eb91a37f111f18583d00cfc.html?fr=income2-wk_app_search_ctrX-search.
|
|
|
[37] |
Chen X J, Wang Y, Li Y F. Removing half antibody byproduct by protein A chromatography during the purification of a bispecific antibody. Protein Expression and Purification, 2020, 172: 105635.
doi: 10.1016/j.pep.2020.105635
|
|
|
[38] |
Wang Y, Chen X J, Wang Y, et al. Removing a difficult-to-separate byproduct by Capto L affinity chromatography during the purification of a WuXiBody-based bispecific antibody. Protein Expression and Purification, 2020, 175: 105713.
doi: 10.1016/j.pep.2020.105713
|
|
|
[39] |
Kluters S, Hafner M, von Hirschheydt T, et al. Solvent modulated linear pH gradient elution for the purification of conventional and bispecific antibodies: Modeling and application. Journal of Chromatography A, 2015, 1418: 119-129.
doi: S0021-9673(15)01359-X
pmid: 26431858
|
|
|
[40] |
Wan Y, Zhang T, Wang Y M, et al. Removing light chain-missing byproducts and aggregates by Capto MMC ImpRes mixed-mode chromatography during the purification of two WuXiBody-based bispecific antibodies. Protein Expression and Purification, 2020, 175: 105712.
|
|
|
[41] |
Wan Y, Wang Y M, Zhang T, et al. Application of pH-salt dual gradient elution in purifying a WuXiBody-based bispecific antibody by MMC ImpRes mixed-mode chromatography. Protein Expression and Purification, 2021, 181: 105822.
doi: 10.1016/j.pep.2021.105822
|
|
|
[42] |
Gagnon P. Improved antibody aggregate removal by hydroxyapatite chromatography in the presence of polyethylene glycol. Journal of Immunological Methods, 2008, 336(2): 222-228.
doi: 10.1016/j.jim.2008.05.002
pmid: 18571666
|
|
|
[43] |
Wang Y, Chen X J, Wang Y, et al. Impact of salt concentration in mobile phase on antibody retention in Protein A, Protein L and KappaSelect affinity chromatography. Protein Expression and Purification, 2021, 178: 105786.
doi: 10.1016/j.pep.2020.105786
|
|
|
[44] |
Chen S W, Tan D, Yang Y S, et al. Investigation of the effect of salt additives in protein L affinity chromatography for the purification of tandem single-chain variable fragment bispecific antibodies. mAbs, 2020, 12(1): 1718440.
|
|
|
[45] |
Hall T, Kelly G M, Emery W R. Use of mobile phase additives for the elution of bispecific and monoclonal antibodies from phenyl based hydrophobic interaction chromatography resins. Journal of Chromatography B, 2018, 1096: 20-30.
doi: S1570-0232(18)30943-7
pmid: 30130673
|
|
|
[46] |
Wollacott R B, Casaz P L, Morin T J, et al. Analytical characterization of a monoclonal antibody therapeutic reveals a three-light chain species that is efficiently removed using hydrophobic interaction chromatography. mAbs, 2013, 5(6): 925-935.
doi: 10.4161/mabs.26192
pmid: 23995619
|
|
|
[47] |
Hall T, Wilson J J, Brownlee T J, et al. Alkaline cation-exchange chromatography for the reduction of aggregate and a mis-formed disulfide variant in a bispecific antibody purification process. Journal of Chromatography B, 2015, 975: 1-8.
doi: 10.1016/j.jchromb.2014.11.002
pmid: 25462105
|
|
|
[48] |
Ishihara T, Miyahara M, Yamamoto K. Monoclonal antibody purification using activated carbon as a replacement for protein A affinity chromatography. Journal of Chromatography B, 2018, 1102-1103: 1-7.
|
|
|
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