Development and application of CK-MB specific monoclonal antibodies

• Published in: Shengwu gongcheng xuebao Vol. 33; no. 1; p. 141
• Main Authors: Chen, Zimin, Zhou, Guoliang, Xu, Weiling, Zheng, Xiaohong, Tong, Xunzhang, Ke, Qishen, Song, Liuwei, Ge, Shengxiang
• Format: Journal Article
• Language: Chinese
• Published: China 25.01.2017
• Subjects: Animals; Antibodies, Monoclonal; Creatine Kinase - immunology; Enzyme-Linked Immunosorbent Assay; Epitopes; Hybridomas; Indicators and Reagents; Isoenzymes; Mice, Inbred BALB C; creatine kinase isoenzyme MB; capture method; acute myocardial infarction (AMI); conformational monoclonal antibodies; monoclonal antibodies
• ISSN: 1000-3061
• DOI: 10.13345/j.cjb.160216

The aim of this study is to develop creatine kinase isoenzyme MB (CK-MB) specific monoclonal antibodies (mAb), and characterize the monoclonal antibody and further development of quantitative detection assay for CK-MB. The BALB/c mice were immunized with purchased CK-MB antigen, then monoclonal antibodies were prepared according to conventional hybridoma technique and screened by indirect and capture ELISA method. To identify the epitopes and evaluate the classification, purchased creatine kinase isoenzyme MB (CK-MM/BB/MB) antigen was used to identify the epitopes, with immunoblotting and synthetic CK-MM and CK-BB in different linear epitope. A double antibody sandwich ELISA was applied to screen the mAb pairs for CK-MB detection, and the quantitative detection assay for CK-MB was developed. We used 74 cases of clinical specimens for comparison of our assay with Roche's CK-MB assay. We successfully developed 22 strains of hybridoms against CK-MB, these mAbs can be divided into linear, partial conformational C