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BRIEF REPORT

Enhanced aggregate separation with Sartobind® Rapid A Protein A membrane in the purification of aggregation-prone antibodies

Gaoya Yuan1 ,  Meng Qu1 ,  Yifeng Li1*
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1 Downstream Process Development (DSPD), WuXi Biologics, Shanghai 200131, China
JBM, e99010089 https://doi.org/10.14440/jbm.0279
Submitted: 28 August 2025 | Revised: 10 October 2025 | Accepted: 13 October 2025 | Published: 10 November 2025
© 2025 by the Author(s). This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution 4.0 International License ( https://creativecommons.org/licenses/by/4.0/ )
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Abstract

Background: Aggregates are common byproducts associated with the production of recombinant antibodies, and their removal poses considerable challenges to the downstream purification. When a Protein A column is used for product capture, large aggregates do not bind due to the size-exclusion effect, whereas small aggregates (e.g., dimers) co-bind with monomers. Although small aggregates bind marginally stronger than the monomer, the difference is usually too small to effect an effective separation of these two species. Thus, Protein A column chromatography generally lacks the ability to separate monomers from co-binding small aggregates. Recently, Protein A membrane has emerged as a promising alternative to resin-based Protein A columns. Objective: This study aimed to evaluate the potential of Sartobind® Rapid A Protein A membrane’s monomer-aggregate separation. Methods: A Protein A column and a Sartobind® Rapid A membrane were used to separately process five culture harvests containing a high percentage of aggregates, and their performances were compared. Aggregate clearance was monitored by analysing relevant elution fractions using size-exclusion chromatography-high-performance liquid chromatography. Results: Sartobind® Rapid A membrane showed stronger aggregate separation capability than the resin-based Protein A column and effectively removed most aggregates in all feed materials. Conclusion: Sartobind® Rapid A membrane outperforms resin-based Protein A columns for antibodies and Fc-fusion proteins with aggregate-rich harvests. By removing most of the aggregates at the capture stage, Sartobind® Rapid A membrane significantly alleviates the purification burden on polishing steps.

Keywords
Aggregate
Antibody
Bispecific antibody
Protein A column
Protein A membrane
Sartobind® Rapid A
Funding
None.
Conflict of interest
Gaoya Yuan, Meng Qu, and Yifeng Li are employees at WuXi Biologics. The company played no role in the study design and the writing of this manuscript.
References
  1. Vázquez-Rey M, Lang DA. Aggregates in monoclonal antibody manufacturing processes. Biotechnol Bioeng. 2011;108(7):1494-1508. doi: 10.1002/bit.23155

 

  1. Li W, Prabakaran P, Chen W, Zhu Z, Feng Y, Dimitrov DS. Antibody aggregation: Insights from sequence and structure. Antibodies (Basel). 2016;5(3):19. doi: 10.3390/antib5030019

 

  1. van der Kant R, Karow-Zwick AR, Van Durme J, et al. Prediction and reduction of the aggregation of monoclonal antibodies. J Mol Biol. 2017;429(8):1244-1261. doi: 10.1016/j.jmb.2017.03.014

 

  1. Li Y. A brief introduction of IgG-like bispecific antibody purification: Methods for removing product-related impurities. Protein Expr Purif. 2019;155:112-119. doi: 10.1016/j.pep.2018.11.011

 

  1. Andrade C, Arnold L, Motabar D, et al. An integrated approach to aggregate control for therapeutic bispecific antibodies using an improved three column Mab platform-like purification process. Biotechnol Prog. 2019;35(1):e2720. doi: 10.1002/btpr.2720

 

  1. Chen SW, Zhang W. Current trends and challenges in the downstream purification of bispecific antibodies. Antib Ther. 2021;4(2):73-88. doi: 10.1093/abt/tbab007

 

  1. Ingavat N, Dzulkiflie N, Liew JM, et al. Investigation on environmental factors contributing to bispecific antibody stability and the reversal of self-associated aggregates. Bioresour Bioprocess. 2024;11(1):82. doi: 10.1186/s40643-024-00796-y

 

  1. Schanzer J, Jekle A, Nezu J, et al. Development of tetravalent, bispecific CCR5 antibodies with antiviral activity against CCR5 monoclonal antibody-resistant HIV-1 strains. Antimicrob Agents Chemother. 2011;55(5):2369-2378. doi: 10.1128/AAC.00215-10

 

  1. Hober S, Nord K, Linhult M. Protein A chromatography for antibody purification. J Chromatogr B Analyt Technol Biomed Life Sci. 2007;848(1):40-47. doi: 10.1016/j.jchromb.2006.09.030

 

  1. Shukla AA, Hubbard B, Tressel T, Guhan S, Low D. Downstream processing of monoclonal antibodies – application of platform approaches. J Chromatogr B Analyt Technol Biomed Life Sci. 2007;848(1):28-39. doi: 10.1016/j.jchromb.2006.09.026

 

  1. Bolton GR, Mehta KK. The role of more than 40 years of improvement in Protein A chromatography in the growth of the therapeutic antibody industry. Biotechnol Prog. 2016;32(5):1193-1202. doi: 10.1002/btpr.2324

 

  1. Yuan G, Qu M, Geng Q, Dong W, Zhang X, Li Y. Antibody aggregation can lead to reduced Protein A binding capacity and low step yield. Protein Expr Purif. 2023;210:106315. doi: 10.1016/j.pep.2023.106315

 

  1. Liu C, Tian M, Dong W, et al. SEC-HPLC analysis of column load and flow-through provides critical understanding of low Protein A step yield. Protein Expr Purif. 2024;216:106418. doi: 10.1016/j.pep.2023.106418

 

  1. Yu D, Song Y, Huang RY, et al. Molecular perspective of antibody aggregates and their adsorption on Protein A resin. J Chromatogr A. 2016;1457:66-75. doi: 10.1016/j.chroma.2016.06.031

 

  1. Zhang Y, Wang Y, Li Y. A method for improving Protein A chromatography’s aggregate removal capability. Protein Expr Purif. 2019;158:65-73. doi: 10.1016/j.pep.2019.02.017

 

  1. Yigzaw Y, Hinckley P, Hewig A, Vedantham G. Ion exchange chromatography of proteins and clearance of aggregates. Curr Pharm Biotechnol. 2009;10(4):421-426. doi: 10.2174/138920109788488842

 

  1. Xu Z, Li J, Zhou JX. Process development for robust removal of aggregates using cation exchange chromatography in monoclonal antibody purification with implementation of quality by design. Prep Biochem Biotechnol. 2012;42(2):183-202. doi: 10.1080/10826068.2012.654572

 

  1. Lu Y, Williamson B, Gillespie R. Recent advancement in application of hydrophobic interaction chromatography for aggregate removal in industrial purification process. Curr Pharm Biotechnol. 2009;10(4):427-433. doi: 10.2174/138920109788488897

 

  1. Gao D, Wang LL, Lin DQ, Yao SJ. Evaluating antibody monomer separation from associated aggregates using mixed-mode chromatography. J Chromatogr A. 2013;1294:70-75. doi: 10.1016/j.chroma.2013.04.018

 

  1. Waller JA, Zheng J, Dyer R, et al. Ceramic hydroxyapatite chromatography plays a critical role in bispecific antibody purification process for impurity removal. Antib Ther. 2022;6(1):30-37. doi: 10.1093/abt/tbac030

 

  1. Ingavat N, Wang X, Liew JM, et al. Harnessing ceramic hydroxyapatite as an effective polishing strategy to remove product- and process-related impurities in bispecific antibody purification. Bioresour Bioprocess. 2023;10(1):93. doi: 10.1186/s40643-023-00713-9

 

  1. Rupčíková V, Molnár T, Kurák T, Polakovič M. Antibody aggregate removal by multimodal chromatography. Molecules. 2025;30(11):2363. doi: 10.3390/molecules30112363

 

  1. Busse RA, Gruenberg M, Kuchemueller KB, Toeppner K, Adametz P, Thom V. Sartobind Rapid A: Comparable high product quality as Protein A resins at high productivity. Application Note; 2022. Available from: https://www. sartorius.hr/media/3u2fwqe3/sartobind-rapid-a-comparable-high-product-quality-as-protein-a-resins-at-high-productivity. pdf [Last accessed on 2025 Oct 01].

 

  1. Grünberg M, Kuchemüller KB, Töppner K, Busse RA. Scalable, robust and highly productive novel convecdiff membrane platform for mAb capture. Membranes. 2022;12(7):677. doi: 10.3390/membranes12070677

 

  1. Osuofa J, Husson SM. Comparative evaluation of commercial Protein A membranes for the Rapid purification of antibodies. Membranes. 2023;13(5):511. doi: 10.3390/membranes13050511

 

  1. Yang S, Braczkowski R, Chen SH, et al. Scalability of Sartobind Rapid A membrane for high productivity monoclonal antibody capture. Membranes. 2023;13(10):815. doi: 10.3390/membranes13100815

 

  1. Gehrmann N, Daxbacher A, Hahn R. Rapid purification of mAb using Protein A membranes yielding high HCP clearance. J Chromatogr B Analyt Technol Biomed Life Sci. 2024;1232:123989. doi: 10.1016/j.jchromb.2023.123989

 

  1. Lu W, Zhang T, Wan Y, Li Y. Protein A membrane shows an advantage over Protein A resin in purifying antibodies/Fc-fusions forming noncovalent aggregates. Chromatogr Sep Tech J. 2024;4:127.

 

  1. Yuan G, Qu M, Li Y. Effective and robust separation of half-antibody byproduct in bispecific antibody purification by Sartobind Rapid A Protein A membrane chromatography. Protein Expr Purif. 2026;237:106823. doi: 10.1016/j.pep.2025.106823
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Journal of Biological Methods, Electronic ISSN: 2326-9901 Print ISSN: TBA, Published by POL Scientific
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