Isolation and characterization of rabies monoclonal antibody charge variants.

Autor: Divase A; Serum Institute of India Pvt. Ltd. Hadapsar, Pune, Maharashtra, India.; Biotechnology Department, Dr. D.Y. Patil Biotechnology and Bioinformatics Institute, Dr. D.Y. Patil Vidyapeeth, Pune, Maharashtra, India., Pisal S; Serum Institute of India Pvt. Ltd. Hadapsar, Pune, Maharashtra, India., Dake MS; Biotechnology Department, Dr. D.Y. Patil Biotechnology and Bioinformatics Institute, Dr. D.Y. Patil Vidyapeeth, Pune, Maharashtra, India., Dakshinamurthy PK; Serum Institute of India Pvt. Ltd. Hadapsar, Pune, Maharashtra, India., Reddy PS; Serum Institute of India Pvt. Ltd. Hadapsar, Pune, Maharashtra, India., Dhere R; Serum Institute of India Pvt. Ltd. Hadapsar, Pune, Maharashtra, India., Kamat C; Serum Institute of India Pvt. Ltd. Hadapsar, Pune, Maharashtra, India., Chahar DS; Serum Institute of India Pvt. Ltd. Hadapsar, Pune, Maharashtra, India., Pal J; Biotechnology Department, Dr. D.Y. Patil Biotechnology and Bioinformatics Institute, Dr. D.Y. Patil Vidyapeeth, Pune, Maharashtra, India., Nawani N; Biotechnology Department, Dr. D.Y. Patil Biotechnology and Bioinformatics Institute, Dr. D.Y. Patil Vidyapeeth, Pune, Maharashtra, India.
Jazyk: angličtina
Zdroj: Electrophoresis [Electrophoresis] 2024 Aug; Vol. 45 (15-16), pp. 1339-1355. Date of Electronic Publication: 2024 May 03.
DOI: 10.1002/elps.202300221
Abstrakt: Current postexposure prophylaxis of rabies includes vaccines, human rabies immunoglobulin (RIG), equine RIG, and recombinant monoclonal antibodies (mAb). In the manufacturing of rabies recombinant mAb, charge variants are the most common source of heterogeneity. Charge variants of rabies mAb were isolated by salt gradient cation exchange chromatography (CEX) to separate acidic and basic and main charge variants. Separated variants were further extensively characterized using orthogonal analytical techniques, which include secondary and tertiary structure determination by far and near ultraviolet circular dichroism spectroscopy. Charge and size heterogeneity were evaluated using CEX, isoelectric focusing (IEF), capillary-IEF, size exclusion chromatography, sodium dodecyl sulfate polyacrylamide gel electrophoresis, and western blotting. Antigen binding affinity was assessed by enzyme linked immuno-sorbent assay and rapid florescence foci inhibition test. Results from structural and physicochemical characterizations concluded that charge variants are formed due to posttranslational modification demonstrating that the charge heterogeneity, these charge variants did neither show any considerable physicochemical change nor affect its biological function. This study shows that charge variants are effective components of mAb and there is no need of deliberate removal, until biological functions of rabies mAb will get affected.
(© 2024 Wiley‐VCH GmbH.)
Databáze: MEDLINE
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