Structural and functional comparability study of anti-CD20 monoclonal antibody with reference product

Sanjay Kumar Singh, Santosh Pokalwar, Sandip Bose, Shivika Gupta, Suhani Almal, Ranjit Sudhakar Ranbhor Sun Pharmaceutical Industries Limited, Tandalja, Vadodara 390 012, India Background: Cell surface protein, CD20, is extensively expressed on the surface of B cells. Antibodies targeting CD20 prote...

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Autores principales:	Singh SK, Pokalwar S, Bose S, Gupta S, Almal S, Ranbhor RS
Formato:	article
Lenguaje:	EN
Publicado:	Dove Medical Press 2018
Materias:	Biosimilar rituximab CD20 monoclonal antibody Medicine (General) R5-920
Acceso en línea:	https://doaj.org/article/b35061115bf340c58a502f433c415c4a
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spelling	oai:doaj.org-article:b35061115bf340c58a502f433c415c4a2021-12-02T03:19:30ZStructural and functional comparability study of anti-CD20 monoclonal antibody with reference product1177-5491https://doaj.org/article/b35061115bf340c58a502f433c415c4a2018-11-01T00:00:00Zhttps://www.dovepress.com/structural-and-functional-comparability-study-of-anti-cd20-monoclonal--peer-reviewed-article-BTThttps://doaj.org/toc/1177-5491Sanjay Kumar Singh, Santosh Pokalwar, Sandip Bose, Shivika Gupta, Suhani Almal, Ranjit Sudhakar Ranbhor Sun Pharmaceutical Industries Limited, Tandalja, Vadodara 390 012, India Background: Cell surface protein, CD20, is extensively expressed on the surface of B cells. Antibodies targeting CD20 protein are being used to treat B-cell malignancies and B-cell mediated autoimmune diseases. Considering the cost of therapy with innovator monoclonal antibodies for these diseases, development of biosimilar products for the treatment of such diseases provides affordable solution to rising healthcare costs. Materials and Methods: Reference products of rituximab (six batches) were procured and stored as per manufacturer's instructions. Cell lines used in bioassay were procured from American Type Culture Collection and all other reagents used for analysis were of analytical grade. Primary structure was studied by intact mass analysis, peptide fingerprinting, peptide mass fingerprinting and sequence coverage analysis. Higher order structure was studied by circular dichroism, ultraviolet-visible spectroscopy, fluorescence spectroscopy, and disulfide bridge analysis. Different isoforms of reference product and SB-02 were identified using capillary isoelectric focusing and capillary zone electrophoresis. Glycosylation was studied by N-glycan mapping using LC-ESI-MS, point of glycosylation, released glycan analysis using ultra performance liquid chromatography (UPLC). Product related impurities such as oligomer content analysis and oxidized impurities were studied using size exclusion chromatography and reverse phase high performance liquid chromatography, respectively.Results and Conclusion: Here, we report physicochemical and biological characterizations of Sun Pharma’s proposed biosimilar (SB-02) to rituximab, a monoclonal anti-CD20 antibody approved for the treatment of non-Hodgkin’s lymphoma and chronic lymphocytic leukemia. SB-02 and rituximab exhibited indistinguishable primary as well as higher-order structure upon analyzing with the array of analytical and extended characterization methods according to statistical methods. The molecule also displayed comparability to reference product in post-translational modifications and charge heterogeneity. In functional bioassays, SB-02 demonstrated comparable potency with respect to reference product. Our results indicate highly similar quality profile between SB-02 and rituximab. Keywords: biosimilar, rituximab, CD20, monoclonal antibody, Biosimilarity, ADCC, CDCSingh SKPokalwar SBose SGupta SAlmal SRanbhor RSDove Medical PressarticleBiosimilarrituximabCD20monoclonal antibodyMedicine (General)R5-920ENBiologics: Targets & Therapy, Vol Volume 12, Pp 159-170 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	Biosimilar rituximab CD20 monoclonal antibody Medicine (General) R5-920
spellingShingle	Biosimilar rituximab CD20 monoclonal antibody Medicine (General) R5-920 Singh SK Pokalwar S Bose S Gupta S Almal S Ranbhor RS Structural and functional comparability study of anti-CD20 monoclonal antibody with reference product
description	Sanjay Kumar Singh, Santosh Pokalwar, Sandip Bose, Shivika Gupta, Suhani Almal, Ranjit Sudhakar Ranbhor Sun Pharmaceutical Industries Limited, Tandalja, Vadodara 390 012, India Background: Cell surface protein, CD20, is extensively expressed on the surface of B cells. Antibodies targeting CD20 protein are being used to treat B-cell malignancies and B-cell mediated autoimmune diseases. Considering the cost of therapy with innovator monoclonal antibodies for these diseases, development of biosimilar products for the treatment of such diseases provides affordable solution to rising healthcare costs. Materials and Methods: Reference products of rituximab (six batches) were procured and stored as per manufacturer's instructions. Cell lines used in bioassay were procured from American Type Culture Collection and all other reagents used for analysis were of analytical grade. Primary structure was studied by intact mass analysis, peptide fingerprinting, peptide mass fingerprinting and sequence coverage analysis. Higher order structure was studied by circular dichroism, ultraviolet-visible spectroscopy, fluorescence spectroscopy, and disulfide bridge analysis. Different isoforms of reference product and SB-02 were identified using capillary isoelectric focusing and capillary zone electrophoresis. Glycosylation was studied by N-glycan mapping using LC-ESI-MS, point of glycosylation, released glycan analysis using ultra performance liquid chromatography (UPLC). Product related impurities such as oligomer content analysis and oxidized impurities were studied using size exclusion chromatography and reverse phase high performance liquid chromatography, respectively.Results and Conclusion: Here, we report physicochemical and biological characterizations of Sun Pharma’s proposed biosimilar (SB-02) to rituximab, a monoclonal anti-CD20 antibody approved for the treatment of non-Hodgkin’s lymphoma and chronic lymphocytic leukemia. SB-02 and rituximab exhibited indistinguishable primary as well as higher-order structure upon analyzing with the array of analytical and extended characterization methods according to statistical methods. The molecule also displayed comparability to reference product in post-translational modifications and charge heterogeneity. In functional bioassays, SB-02 demonstrated comparable potency with respect to reference product. Our results indicate highly similar quality profile between SB-02 and rituximab. Keywords: biosimilar, rituximab, CD20, monoclonal antibody, Biosimilarity, ADCC, CDC
format	article
author	Singh SK Pokalwar S Bose S Gupta S Almal S Ranbhor RS
author_facet	Singh SK Pokalwar S Bose S Gupta S Almal S Ranbhor RS
author_sort	Singh SK
title	Structural and functional comparability study of anti-CD20 monoclonal antibody with reference product
title_short	Structural and functional comparability study of anti-CD20 monoclonal antibody with reference product
title_full	Structural and functional comparability study of anti-CD20 monoclonal antibody with reference product
title_fullStr	Structural and functional comparability study of anti-CD20 monoclonal antibody with reference product
title_full_unstemmed	Structural and functional comparability study of anti-CD20 monoclonal antibody with reference product
title_sort	structural and functional comparability study of anti-cd20 monoclonal antibody with reference product
publisher	Dove Medical Press
publishDate	2018
url	https://doaj.org/article/b35061115bf340c58a502f433c415c4a
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Structural and functional comparability study of anti-CD20 monoclonal antibody with reference product

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