铜离子鳌合物人工抗原的合成与鉴定

Synthesis and Identification of Artificial Antigen Against Copper Ionic Chelate

该研究为获得铜离子螯合物人工抗原,采用异硫氰酯法,以异硫氰酸苄基乙二胺四乙酸(ITCBE)为双功能螯合剂,将载体蛋白牛血清白蛋白(BSA)和卵清蛋白(OVA)分别与Cu^(2+)偶联,得到人工抗原(Cu^(2+)-ITCBE-BSA)和包被原(Cu^(2+)-ITCBE-OVA)。通过紫外分光光度法(UV)和电感耦合等离子体原子发射光谱法(ICP-AES),以及变性聚丙烯凝胶电泳(SDS-PAGE)和免疫学检测(Immunoassay)的方法对所获得的免疫原进行鉴定。结果显示获得人工抗原的浓度为9.21 mg/m L;Cu^(2+)的含量为16.83 g/mL;免疫小鼠间接ELISA效价达1:51 200以上,阻断ELISA检测Cu^(2+)-EDTA的半数抑制浓度(IC_(50))为36.73 ng/mL,为抗铜离子单克隆抗体(mAb)的进一步研制奠定了基础。

To obtain the artificial immunogen against copper ionic chelate. Copper ion [Cu（II）]was coupled to bovine serum albumin（BSA） and ovalbumin（OVA） which were carrier proteins via a bifunctional chelating agent Isothiocyanobenzyl-EDTA（ITCBE）. The immunogen was identified by ultraviolet spectrophotometry（UV）,inductively coupled plasma atomic emission spectrometry（ICP-AES）,denaturing polyacrylamide gel electrophoresis（SDS-PAGE）,and immunoassay. The results showed that the concentration of artificial antigen and Cu^2＋ were 9.21 mg/mL and 16.83 μg/m L. The Cu^2＋-EDTA p Ab of mouses had the titers of 1：51200 by indirect ELISA and the IC_（50） of Cu^2＋-EDTA is 36.73 ng/m L by blocking ELISA. The artificial antigen was successfully prepared and has a good immunogenicity,which provided foundations for the development of m Ab against copper ions.