Prokaryotic Expression And Establishment Of Multiple Immunoassays For Human ST2 Protein

In recent years,the number of people with myocardial injury has been increasing.Human soluble growth stimulative expression gene 2 protein(ST2)is a new marker of myocardial injury,which is of guiding significance for the detection of treatment of acute myocardial infarction(AMI)and heart failure(HF)patients.Multiple immunoassay methods for studying ST2 protein provide laboratory basis for the development of clinical application and kit.In this paper,the expression vector pET-15b-ST2 was constructed to be expressed in E.coli BL21(DE3).Expression conditions were selected for induction with 1 mM IPTG for 24 h at 16°C,110 rpm.After purification,the purity of soluble ST2 protein was as high as 95% and the concentration was 6.25 mg/ml.In this study,we established a variety of immunoassay methods for ST2 protein,including enzyme-linked immunosorbent assay,chemiluminescence method based on magnetic microspheres,colloidal gold immunoassay.After a series of optimization of experimental conditions,the best process was established.The enzyme-linked immunosorbent assay established in this study was compared with the commercially available ELISA method.The method has good consistency and is applied to the detection of serum soluble ST2 in clinical samples with good results.