简介：Objective:Toachieveanoptimizedmethodforsolubleexpressionofhumancarboxylesterase1(hCE-1)inescherichiacoilandpurificationbyNi2+-NTAagaroseaffinitychromatography,togetimprovedproteinyieldandpurityforfurtherdevelopmentofhepatocellularcarcinoma(HCC)diagnosisELISAkits.Methods:ThebestantigenepitopesofhCE1werepredictedbycomparingsecondarystructure,flexibleregions,hydrophilicity,antigenicindexsurfaceprobabilityofresidues.Afterwards,pET-42a(+)withaHis-tagandaGST-tagwasappliedtoformrecombinantplasmidpET-42a(+)/hCE1,whichfacilitatedpurificationwhenusingNi2+-NTAagaroseaffinitychromatography.ProteinqualitywasmeasuredbySDS-PAGEandBCAproteinassay.Western-blotidentificationwasalsoperformedtoensurethecorrectexpressionofhCE1protein.Results:Theresiduesfrom500to567nearC-terminalofhCE1proteinwereconsideredthebestepitopeswhichexhibitedhighhydrophilicityandhighsurfaceprobabilityandrelativelyflexiblesecondarystructureandlowhomologycomparedwithhCE2andhCE3.His-hCE1500-567fusionproteinwasachievedbyIPTG-inductedexpressionwithanexpectedmassof42kDa.Afterpurification,thefinalproductwasspeciallyidentified,whichreachedover95%purityandmorethan10mg/Lofmicrobialculture.InWesternblot,thepurifiedfusionproteinwasrecognizedbyanti-hCE1monoclonalantibody,alongwithprevioussequencingvalidation,whichdemonstratedthecorrectpreparationofsolublehCE1protein.Conclusion:ThisisanefficaciousandaffordablestrategytogeneratefusionhCE1ofhighqualityinEcoli,whichfacilitatespreparationofhCE1monoclonalantibodyandfurtherHCCdiagnosisresearch.

简介：胰岛移植是治疗1型糖尿病的有效方法,但是胰岛供体短缺、微环境破坏、免疫排斥、营养供应等是胰岛移植中的难题和挑战。组织工程方法是用支架材料或细胞与胰岛共移植,改善胰岛的存活和功能。而分子影像技术为移植后胰岛的生物学过程提供了有效的检测手段。文章对近年来组织工程技术改善胰岛移植的生存微环境干细胞治疗,以及分子影像方法进行胰岛移植后活体、实时、动态观测的研究进行了综述,对目前胰岛移植存在的问题和未来的发展方向进行了讨论。