1.Cloning,Expression and Diagnostic Application of Different Fragments of Human Carboxypeptidase
Jiwen HU ; Xiqin YANG ; Keliang ZONG ; Lanqin GUO ; Xiaoguo SONG ; Guohua WANG ; Ximing LIU ; Cuixia ZHU ; Yanfeng ZHAO ; Xiaoyan FENG ; Heqiu ZHANG
Journal of Modern Laboratory Medicine 2015;(4):10-13
Objective To obtain different fragments of human carboxypeptidase H,and evaluate the diagnostic application of the recombination carboxypeptidase H in detecting autoantibody.Methods The coding gene of carboxypeptidase H was ob-tained by RT-PCR.The corresponding prokaryotic expression vectors were constructed and transformed into E.coli to in-duce the expression of the recombination different fragments of carboxypeptidase H.Using these antigen fragments as the coating antigens,the enzyme-linked immunosorbent assay (ELISA)was established for the detection of carboxypeptidase H autoantibody in 95 newly diagnosed type 2 diabetes patients.Results Three fragments of human carboxypeptidase H were obtained,in which the 42~476aa fragment antigen was ideal one.Using the full-length carboxypeptidase H as coating anti-gen,the positive rate of carboxypeptidase H autoantibody was 8.42%.Conclusion Because of the favorable antigenicity,the 42~476aa fragment antigen of carboxypeptidase H could be the candidate antigen for discrimination and diagnosis of latent autoimmune diabetes in adults.
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