Generation of a human monoclonal antibody to cross-reactive material 197 (CRM197) and development of a sandwich ELISA for CRM197 conjugate vaccines
| Autor: | Jimin Wi, Dain Kim, Heesu Chae, Chankyu Lee, Heeyoun Kim, Jun-Yeol Yoon, Sang Kyu Kim, Hyo Jeong Hong, Gyunghee Jo, Se-Ho Kim, Hyeseon Yoon |
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| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Diphtheria toxin Phage display biology Chemistry medicine.drug_class General Medicine Monoclonal antibody Applied Microbiology and Biotechnology Molecular biology 03 medical and health sciences 030104 developmental biology Immune system Antigen Polyclonal antibodies biology.protein medicine Antibody Biotechnology Conjugate |
| Zdroj: | Journal of Microbiology and Biotechnology. 28:2113-2120 |
| ISSN: | 1738-8872 1017-7825 |
| DOI: | 10.4014/jmb.1810.10009 |
| Popis: | Cross-reactive material 197 (CRM₁₉₇) is a non-toxic mutant of diphtheria toxin containing a single amino acid substitution of glycine 52 with glutamic acid. CRM₁₉₇ has been used as a carrier protein for poorly immunogenic polysaccharide antigens to improve immune responses. In this study, to develop a sandwich ELISA that can detect CRM₁₉₇ and CRM₁₉₇ conjugate vaccines, we generated a human anti-CRM₁₉₇ monoclonal antibody (mAb) 3F9 using a phage-displayed human synthetic Fab library and produced mouse anti-CRM₁₉₇ polyclonal antibody. The affinity (KD) of 3F9 for CRM₁₉₇ was 3.55 nM, based on Bio-Layer interferometry, and it bound specifically to the B fragment of CRM₁₉₇. The sandwich ELISA was carried out using 3F9 as a capture antibody and the mouse polyclonal antibody as a detection antibody. The detection limit of the sandwich ELISA was ＜1 ng/ml CRM₁₉₇. In addition, the 3F9 antibody bound to the CRM₁₉₇-polysaccharide conjugates tested in a dose-dependent manner. This ELISA system will be useful for the quantification and characterization of CRM₁₉₇ and CRM₁₉₇ conjugate vaccines. To our knowledge, this study is the first to generate a human monoclonal antibody against CRM₁₉₇ and to develop a sandwich ELISA for CRM₁₉₇ conjugate vaccines. |
| Databáze: | OpenAIRE |
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