Expression And Application Of17β-hydroxysteroid Dehydrogenase Type10

17β-hydroxysteroid dehydrogenase type10is a multifunctional emyze which can catalyse various substrates. And it’s confimed so far that the emyze involved in the metabolism of3-hydroxyacyl-CoA, isoleucine, sex steroid hormones and neuroactive steroids. Specifically,17β-HSD10takes part in cognitive activities of the brain. More over, substantive17β-HSD10can be detected in the brain of human or mouse who illed with Alzheimer’s disease. It is indicated that the emyze might be developed to be a kind of biomarker to diagnose AD.RT-PCR was employed to get the gene of17β-HSD10of mouse liver, and a prokaryotic protein expression system pET15b-17β-HSD10/E.coli BL21(DE3) which induced with isopropyl-1-thio-β-galactopy ranoside(IPTG) for recombinant protein expression was constructed subsequently. The target protein was purified using His-Binding-resin column, and immunized to BALB/c mice and rabbits, serum total IgGs from immunized animals were purified by ammonium sulfate precipitation method. Two antibodies we prepared previously were used when we established a Double-antibody Sandwich ELISA about17β-HSD10. Finally we got the concentration of1.5mg/mL of17β-HSD10protein with molecular weight of29.5kDa, and polyclonal antibodies from mouse and rabbit with the titer of1.25x104and2.5x104respectively. The concentration of0.1μ,g/mL of17β-HSD10can be detected by the Double-antibody Sandwich ELISA we established, and the assay was proved to be with good sensitivity and specificity. Our subject will be helpful for clinical diagnosis of Alzheimer’s disease.