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Quantitation and Identification of Therapeutic Anti-CD22 Monoclonal Antibodies in a Cell-Based ELISA Method

Since they lack native soluble membrane antigens, the analysis and selection of antigen-specific antibodies are commonly performed on whole live cells. Here, we have developed a simple and convenient enzyme-linked immunosorbent assay (ELISA) based on cell membrane antigens. Soluble cell membrane pro...

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Detalles Bibliográficos
Autores principales: Fu, Shengyu, Zhao, Qi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9396980/
https://www.ncbi.nlm.nih.gov/pubmed/35997347
http://dx.doi.org/10.3390/antib11030053
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author Fu, Shengyu
Zhao, Qi
author_facet Fu, Shengyu
Zhao, Qi
author_sort Fu, Shengyu
collection PubMed
description Since they lack native soluble membrane antigens, the analysis and selection of antigen-specific antibodies are commonly performed on whole live cells. Here, we have developed a simple and convenient enzyme-linked immunosorbent assay (ELISA) based on cell membrane antigens. Soluble cell membrane proteins isolated from Raji cells were immobilized on the polystyrene microplate, which permitted the assessment of a therapeutic anti-CD22 monoclonal antibody. The experiments showed less variability in the intra-assay. Compared to the living cell ELISAs, the advantage of the assay is avoiding cell losses and high variation of optical density (OD) readings. We provide a quantitative and reproducible ELISA that can be potentially applied to the development of specific antibodies against cell surface antigens.
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spelling pubmed-93969802022-08-24 Quantitation and Identification of Therapeutic Anti-CD22 Monoclonal Antibodies in a Cell-Based ELISA Method Fu, Shengyu Zhao, Qi Antibodies (Basel) Article Since they lack native soluble membrane antigens, the analysis and selection of antigen-specific antibodies are commonly performed on whole live cells. Here, we have developed a simple and convenient enzyme-linked immunosorbent assay (ELISA) based on cell membrane antigens. Soluble cell membrane proteins isolated from Raji cells were immobilized on the polystyrene microplate, which permitted the assessment of a therapeutic anti-CD22 monoclonal antibody. The experiments showed less variability in the intra-assay. Compared to the living cell ELISAs, the advantage of the assay is avoiding cell losses and high variation of optical density (OD) readings. We provide a quantitative and reproducible ELISA that can be potentially applied to the development of specific antibodies against cell surface antigens. MDPI 2022-08-16 /pmc/articles/PMC9396980/ /pubmed/35997347 http://dx.doi.org/10.3390/antib11030053 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Fu, Shengyu
Zhao, Qi
Quantitation and Identification of Therapeutic Anti-CD22 Monoclonal Antibodies in a Cell-Based ELISA Method
title Quantitation and Identification of Therapeutic Anti-CD22 Monoclonal Antibodies in a Cell-Based ELISA Method
title_full Quantitation and Identification of Therapeutic Anti-CD22 Monoclonal Antibodies in a Cell-Based ELISA Method
title_fullStr Quantitation and Identification of Therapeutic Anti-CD22 Monoclonal Antibodies in a Cell-Based ELISA Method
title_full_unstemmed Quantitation and Identification of Therapeutic Anti-CD22 Monoclonal Antibodies in a Cell-Based ELISA Method
title_short Quantitation and Identification of Therapeutic Anti-CD22 Monoclonal Antibodies in a Cell-Based ELISA Method
title_sort quantitation and identification of therapeutic anti-cd22 monoclonal antibodies in a cell-based elisa method
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9396980/
https://www.ncbi.nlm.nih.gov/pubmed/35997347
http://dx.doi.org/10.3390/antib11030053
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