Cross-reactive material 197 (CRM<sub>197</sub>) is a non-toxic mutant of diphtheria toxin containing a single amino acid substitution of glycine 52 with glutamic acid. CRM<sub>197</sub> has been used as a carrier protein for po...
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https://www.riss.kr/link?id=A106006808
2018
-
SCIE,SCOPUS,KCI등재
학술저널
2113-2120(8쪽)
0
상세조회0
다운로드다국어 초록 (Multilingual Abstract)
Cross-reactive material 197 (CRM<sub>197</sub>) is a non-toxic mutant of diphtheria toxin containing a single amino acid substitution of glycine 52 with glutamic acid. CRM<sub>197</sub> has been used as a carrier protein for po...
Cross-reactive material 197 (CRM<sub>197</sub>) is a non-toxic mutant of diphtheria toxin containing a single amino acid substitution of glycine 52 with glutamic acid. CRM<sub>197</sub> 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 CRM<sub>197</sub> and CRM<sub>197</sub> conjugate vaccines, we generated a human anti-CRM<sub>197</sub> monoclonal antibody (mAb) 3F9 using a phage-displayed human synthetic Fab library and produced mouse anti-CRM197 polyclonal antibody. The affinity (K<sub>D</sub>) of 3F9 for CRM<sub>197</sub> was 3.55 nM, based on Bio-Layer interferometry, and it bound specifically to the B fragment of CRM<sub>197</sub>. 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 CRM<sub>197</sub>. In addition, the 3F9 antibody bound to the CRM<sub>197</sub>-polysaccharide conjugates tested in a dose-dependent manner. This ELISA system will be useful for the quantification and characterization of CRM<sub>197</sub> and CRM<sub>197</sub> 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 CRM<sub>197</sub> conjugate vaccines.
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