分泌抗心脏肌凝蛋白及其轻链单克隆抗体杂交瘤细胞系的建立及初步鉴定

心脏肌凝蛋白(Cardiac myosin,CM)及其轻链(CM—LC)是心肌特有的收缩蛋白质,其化学结构和免疫学性质均与其它组织来源者不同。当急性心肌梗塞(AMI)造成心肌细胞缺血坏死时,血清CM—LC水平升高。故可用于临床诊断AMI。这一诊断指标的特异性、敏感性、与梗塞灶大小

HYBRIDOMA CELL LINES SECRETING MONOC-LONAL ANTIBODIES AGAINST CARDIAC MYOSIN AND ITS LIGHT CHAINS

Four mouse hybridoma cell lines secreting monoclonal antibodies (McAb) against human and canine cardiac myosins (CM) and their light chains (CM-LC) were produced. Spleen cells of BALB/c mice immunized with CM antigens from human or canine ventricular muscle were fused with NS-1 or SP2/0 mouse myeloma cell lines. The specific antibody positive wells were detected by ELISA using horse-radish peroxidase-rab-bit anti-mouse Ig sera (HRP-RAM) as the labeled antibody. After four or five subclonings, cell lines FM1 (3E3), FM2 (2ElO), FM3 (1F3) and FM4 (1A4), grew well in culture bottles and could produce mouse antibody-enriched ascites. Results of testing the four McAbs with cardiac-, skeletal- and smooth-muscle myosins or their subunits from human and canine sources by ELISA showed that McAb FMl (IgG1) was human myosin heavy chain specific; and FM3 (IgM), CM-LC specific. McAbs FM2 and FM4 (both IgM) were myosin heavy chain specific and had complete cross reaction with cardiac and skeletal muscle myosins. It is indicated that specific McAbs could be used to detect serum CM-LC and image myocardial infarct for the clinical diagnosis of acute myocardial infarction.