Studies on affinity of cTnⅠ monoclonal antibody with different epitopes to cTnⅠ-TnC complex
10.3969/j.issn.1000-484X.2025.01.028
- VernacularTitle:不同抗原表位的cTnⅠ单克隆抗体对cTnⅠ-TnC复合物亲和性的研究
- Author:
Chengshan HE
1
;
Yang LIU
;
Luyao LIU
;
Yu WU
;
Zhicheng LU
Author Information
1. 上海中医药大学附属第七人民医院医学检验科,上海 200137
- Publication Type:Journal Article
- Keywords:
Cardiac troponin Ⅰ;
Troponin C;
Monoclonal antibody;
Affinity
- From:
Chinese Journal of Immunology
2025;41(1):189-194
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate affinity of monoclonal antibody(MAb)of different cardiac troponinⅠ(cTnⅠ)epi-topes to cTnⅠ-troponin C(cTnⅠ-TnC)antigen,and to screen novel cTnⅠ-specific antibodies for clinical detection kit development.Methods:Based on technology platform of microarray chemiluminescence immunoassay,cTnⅠ-TnC antigen spot sample hard matrix chip was used,mouse derived cTnⅠ MAb was used as antibody to be detected,HRP-sheep-anti-mouse-IgG was used as antibody to detect,and experimental conditions were optimized for anti-origin solution,antigen spot sample concentration and antibody to be detected concentration.Affinity of cTnⅠ MAb with different antigenic epitopes to cTnⅠ-TnC was detected by competition method.Graded concentrations of cTnⅠ-TnC were added to experimental group,and diluents of equal volume were added to control group.Reaction signals of different cTnⅠ MAb were detected and affinity constants were calculated.Results:Antigen spot sample liquid was P105,antigen spot sample concentration was 0.1 mg/ml,and antibody concentration to be detected was 80 ng/ml were optimal experi-mental conditions.20c6cc and 7B9cc MAb containing TnC antibodies had the best affinity,and 20c6cc MAb reached 3.18×106 L/mol.cTnⅠ MAb located at C-end of peptide chain in a.a.r 130～145,169～178 and 190～196 regions showed good affinity,among which MAb 625(a.a.r 169～178)was 6.37×105 L/mol,showing the strongest affinity among cTnⅠ MAb.Conclusion:MAb has good affinity with cTnⅠ-TnC antigen at C-end of cTnⅠ peptide chain a.a.r 130～145,169～178,190～196 and TnC region.Designing a new type of cTnⅠ complement antibody targeting this region can be a new way to solve negative interference of autoantibody and peptide proteolysis.
