earticle

영어

The cardiac troponin I (cTnI), one of biomarkers to diagnose acute myocardial infarction. In order to produce recombinant single chain variable fragments (ScFv) antibody against cTnI, Single-stranded complementary DNAs were synthesized from the hybridoma cell lines #6 and #12 which produce specific antibodies against cTnI. Heavy chain variable region and light chain variable region were amplified by polymerase chain reaction. Each amplified gene was linked using (Gly4Ser)3 linker. Then each scFv gene fragment was cloned into pET26b vector containing signal sequence pelB for secretion of target proteins. ELASA analysis revealed that the secreted scFvs were active against cTnI. In order to display on the surface of M18 phage display the cTnI-ScFv12 gene fragment fused with PⅢ gene derived from M18mp19 was cloned into pUC119 phagemid vector. ScFv-cTnI12 was successfully displayed on M13 phage, and antibody engineering will be progress to improve binding affinity of the antibody.