{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,27]],"date-time":"2026-05-27T21:45:10Z","timestamp":1779918310000,"version":"3.53.1"},"reference-count":35,"publisher":"MDPI AG","issue":"10","license":[{"start":{"date-parts":[[2025,10,1]],"date-time":"2025-10-01T00:00:00Z","timestamp":1759276800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["www.mdpi.com"],"crossmark-restriction":true},"short-container-title":["Membranes"],"abstract":"<jats:p>In the production of recombinant antibody\/Fc-fusion proteins using mammalian cells, many aggregates often form alongside the target proteins, particularly with bispecific antibodies. To ensure the safety of biological products, it is essential to control the amount of aggregates within a specific range. A traditional downstream process typically involves using Protein A (ProA) resin to capture the target antibody, followed by two polishing steps to ensure purity; for instance, using an anion exchange chromatography (AEX) in flow-through mode and a cation exchange chromatography (CEX) in binding\u2013elution mode. In this study, we choose a Dual Action Fab (DAF), which can bind two antigens and is prone to aggregation when expression in CHO (Chinese Hamster Ovary) cells. We introduce hydrophobic interaction membrane chromatography (HIMC) operating in flow-through mode, which enhances production efficiency while reducing costs and the risks associated with column packing. We evaluated the impact of the operating buffer system, as well as the pH and conductivity of the loading samples, on aggregate removal using HIMC. Additionally, we investigated the mechanism of aggregate binding and found that loading conditions had a limited impact on this process. Overall, our findings indicate that employing HIMC can achieve a 20% reduction in aggregate levels. These results demonstrate that HIMC in flow-through mode is an effective and robust approach for reducing aggregates during antibody purification.<\/jats:p>","DOI":"10.3390\/membranes15100299","type":"journal-article","created":{"date-parts":[[2025,10,1]],"date-time":"2025-10-01T07:58:13Z","timestamp":1759305493000},"page":"299","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Removal of Aggregates During Bispecific Antibody Purification Using Hydrophobic Interaction Chromatography"],"prefix":"10.3390","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0009-0007-0555-8758","authenticated-orcid":false,"given":"Puya","family":"Zhao","sequence":"first","affiliation":[{"name":"Shanghai AsymChem Biotechnology Co., Ltd., Building 8, No.12, Lane 855, Jinzheng Road, Jinshan Industrial Park, Shanghai 201507, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yue","family":"Qi","sequence":"additional","affiliation":[{"name":"Shanghai AsymChem Biotechnology Co., Ltd., Building 8, No.12, Lane 855, Jinzheng Road, Jinshan Industrial Park, Shanghai 201507, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kai","family":"Gao","sequence":"additional","affiliation":[{"name":"Shanghai AsymChem Biotechnology Co., Ltd., Building 8, No.12, Lane 855, Jinzheng Road, Jinshan Industrial Park, Shanghai 201507, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,10,1]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"105647","DOI":"10.1016\/j.pep.2020.105647","article-title":"A potential downstream platform approach for WuXiBody-based IgG-like bispecific antibodies","volume":"173","author":"Guo","year":"2020","journal-title":"Protein Expr. 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