Washing with alkaline solutions in protein A purification improves physicochemical properties of monoclonal antibodies

Abstract Protein A affinity chromatography has been widely used for both laboratory scale purification and commercial manufacturing of monoclonal antibodies and Fc-fusion proteins. Protein A purification is specific and efficient. However, there still remain several issues to be addressed, such as i...

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Autores principales:	Yuichi Imura, Toshiaki Tagawa, Yuya Miyamoto, Satoshi Nonoyama, Hiroshi Sumichika, Yasuhiro Fujino, Masaya Yamanouchi, Hideo Miki
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/0ca246364b864224b7da37a28cf7a642
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spelling	oai:doaj.org-article:0ca246364b864224b7da37a28cf7a6422021-12-02T10:49:29ZWashing with alkaline solutions in protein A purification improves physicochemical properties of monoclonal antibodies10.1038/s41598-021-81366-62045-2322https://doaj.org/article/0ca246364b864224b7da37a28cf7a6422021-01-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-81366-6https://doaj.org/toc/2045-2322Abstract Protein A affinity chromatography has been widely used for both laboratory scale purification and commercial manufacturing of monoclonal antibodies and Fc-fusion proteins. Protein A purification is specific and efficient. However, there still remain several issues to be addressed, such as incomplete clearance of impurities including host cell proteins, DNA, aggregates, etc. In addition, the effects of wash buffers in protein A purification on the physicochemical characteristics of antibodies have yet to be fully understood. Here we found a new purification protocol for monoclonal antibodies that can improve physicochemical properties of monoclonal antibodies simply by inserting an additional wash step with a basic buffer after the capture step to the conventional protein A purification. The effects of the alkaline wash on monoclonal antibodies were investigated in terms of physicochemical characteristics, yields, and impurity clearance. The simple insertion of an alkaline wash step resulted in protection of antibodies from irreversible aggregation, reduction in free thiols and impurities, an improvement in colloidal and storage stability, and enhanced yields. This new procedure is widely applicable to protein A affinity chromatography of monoclonal antibodies.Yuichi ImuraToshiaki TagawaYuya MiyamotoSatoshi NonoyamaHiroshi SumichikaYasuhiro FujinoMasaya YamanouchiHideo MikiNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-11 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Yuichi Imura Toshiaki Tagawa Yuya Miyamoto Satoshi Nonoyama Hiroshi Sumichika Yasuhiro Fujino Masaya Yamanouchi Hideo Miki Washing with alkaline solutions in protein A purification improves physicochemical properties of monoclonal antibodies
description	Abstract Protein A affinity chromatography has been widely used for both laboratory scale purification and commercial manufacturing of monoclonal antibodies and Fc-fusion proteins. Protein A purification is specific and efficient. However, there still remain several issues to be addressed, such as incomplete clearance of impurities including host cell proteins, DNA, aggregates, etc. In addition, the effects of wash buffers in protein A purification on the physicochemical characteristics of antibodies have yet to be fully understood. Here we found a new purification protocol for monoclonal antibodies that can improve physicochemical properties of monoclonal antibodies simply by inserting an additional wash step with a basic buffer after the capture step to the conventional protein A purification. The effects of the alkaline wash on monoclonal antibodies were investigated in terms of physicochemical characteristics, yields, and impurity clearance. The simple insertion of an alkaline wash step resulted in protection of antibodies from irreversible aggregation, reduction in free thiols and impurities, an improvement in colloidal and storage stability, and enhanced yields. This new procedure is widely applicable to protein A affinity chromatography of monoclonal antibodies.
format	article
author	Yuichi Imura Toshiaki Tagawa Yuya Miyamoto Satoshi Nonoyama Hiroshi Sumichika Yasuhiro Fujino Masaya Yamanouchi Hideo Miki
author_facet	Yuichi Imura Toshiaki Tagawa Yuya Miyamoto Satoshi Nonoyama Hiroshi Sumichika Yasuhiro Fujino Masaya Yamanouchi Hideo Miki
author_sort	Yuichi Imura
title	Washing with alkaline solutions in protein A purification improves physicochemical properties of monoclonal antibodies
title_short	Washing with alkaline solutions in protein A purification improves physicochemical properties of monoclonal antibodies
title_full	Washing with alkaline solutions in protein A purification improves physicochemical properties of monoclonal antibodies
title_fullStr	Washing with alkaline solutions in protein A purification improves physicochemical properties of monoclonal antibodies
title_full_unstemmed	Washing with alkaline solutions in protein A purification improves physicochemical properties of monoclonal antibodies
title_sort	washing with alkaline solutions in protein a purification improves physicochemical properties of monoclonal antibodies
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/0ca246364b864224b7da37a28cf7a642
work_keys_str_mv	AT yuichiimura washingwithalkalinesolutionsinproteinapurificationimprovesphysicochemicalpropertiesofmonoclonalantibodies AT toshiakitagawa washingwithalkalinesolutionsinproteinapurificationimprovesphysicochemicalpropertiesofmonoclonalantibodies AT yuyamiyamoto washingwithalkalinesolutionsinproteinapurificationimprovesphysicochemicalpropertiesofmonoclonalantibodies AT satoshinonoyama washingwithalkalinesolutionsinproteinapurificationimprovesphysicochemicalpropertiesofmonoclonalantibodies AT hiroshisumichika washingwithalkalinesolutionsinproteinapurificationimprovesphysicochemicalpropertiesofmonoclonalantibodies AT yasuhirofujino washingwithalkalinesolutionsinproteinapurificationimprovesphysicochemicalpropertiesofmonoclonalantibodies AT masayayamanouchi washingwithalkalinesolutionsinproteinapurificationimprovesphysicochemicalpropertiesofmonoclonalantibodies AT hideomiki washingwithalkalinesolutionsinproteinapurificationimprovesphysicochemicalpropertiesofmonoclonalantibodies
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Washing with alkaline solutions in protein A purification improves physicochemical properties of monoclonal antibodies

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