关键词: 肌酸激酶同工酶 CK-MB, 急性心肌梗死, 基因工程抗体

Abstract: Objective Construct heterodimeric CK-MB recombinant protein and prepare its specific geneengineered antibody as the diagnostic raw material of CK-MB quantitative detection kit, so as to assist theearly diagnosis of acute myocardial infarction in clinic, and to improve the accuracy of diagnosis. MethodMouse plasma cells were obtained by obtaining heterodimeric CK-MB through co-expression vector andanimal immunization, further, single mouse B cells specific for antigen were isolated using single-cellphotoconductive system, and genetically engineered antibodies were constructed after obtaining the genesequences of the variable regions of the light and heavy chains of the antibodies through single-cell PCR.Candidate antibodies were screened based on preliminary functional validation, and the optimal pairingscheme was further explored by checkerboard titration, and a CK-MB quantitative detection kit wasinitially assembled using the principle of double-antibody sandwich ELISA. Result CK-MB immunogenwas successfully obtained; single plasma cells specific for CK-MB antigen were screened by single B-cellphotoconductivity and two strains of mouse-derived genetically engineered antibodies were prepared, andboth of them had the ability of recognizing the natural CK-MB; two pairwise protocols were screened by double-antibody sandwich ELISA, and the R2of the fitted curves were both greater than 0. 99.Conclusion Mouse-derived genetically engineered antibody targeting CK-MB was successfully prepared,and the specificity and affinity of the antibody was preliminarily verified by ELISA, laying a solidfoundation for the further development of a new type of CK-MB quantitative detection kit with highspecificity and high sensitivity.

Key words: creatine kinase isoenzyme CK-MB, acute myocardial infarction, geneticallyengineered antibody