人雌激素受体BCD区段在大肠杆菌的高效表达及其抗体于乳腺癌石蜡切片免疫组化应用

OVERPRODUCTION OF BCD DOMAIN OF HUMAN ESTROGEN RECEPTOR IN ESCHERICHIA COLI AND ITS ANTISERUM CHARACTERIZATION AND APPLICATION

目的：获取人雌激素受体（ＥＲ）及其抗体。方法：扩增编码ＥＲＢＣＤ区的基因，在大肠杆菌中表达ＭＳ２－ＥＲ融合蛋白。免疫动物，得抗ＥＲ血清，以此检测５０余例乳腺良、恶性肿瘤的ＥＲ表达，并与经典检测ＥＲ的ＤＣＣ法及进口抗ＥＲ单克隆抗体Ｃ－３１４结果相比较。结果：ＭＳ２－ＥＲ融合蛋白有天然ＥＲ。结构，其抗血清特异针对野生型ＥＲ，其ＩＨＣ结果与ＤＣＣ及Ｃ－３１４抗ＥＲ单克隆抗体有高符合率，既具备ＩＨＣ方法的特点，又免去需微波处理这一步骤。结论：自制抗ＥＲ血清可用来检测ＥＲ表达，为进一步获取其单克隆抗体及推广奠定基础。

Objective:To gain the human estrogen receptor(ER)and its antiserum.Methods:We have amplified a gene fragment encoding the BCD domain of ER and overproduced MS2-ER fusion protein in E.coli,By immunization with the purified MS2-ER, a serum to the ER was raised.And with this antiserum,we not only examined the ER expression in fifties breast cancers and benign tumors,but also contrast this to the results of DCC and IHC of the McAb to ER,C-314,Results:The MS2-ER fusion protein has native constructive of ER,The antiserum we have gained exclusively recognizes the ER and there is high correspondence among the three results:our antiserum,C-314 and DCC.Conclusion:IHC of the antiserum we have gained would serve as a powerful tool to detect and analyse the ER in paraffin embedded section.