重金属镉离子免疫抗原初步制备与鉴定

Primary preparation and identification of immune antigen chelating heavy metal ion cadmium

选用螫合剂Isothiocyanobenzyl-EDTA（IEDTA）分别桥接载体蛋白KLH和BSA。络合重金属镉离子，初步制备免疫抗原和检测抗原，免疫Balb／C小鼠并制备抗血清；利用紫外分光光度法、三硝基苯磺酸法（TNBs）和测定抗体效价初步鉴定抗原。建立了重金属镉离子ELISA快速检测技术及单克隆抗体高效特异的免疫与检测抗原。抗原紫外吸收峰与载体蛋白紫外吸收峰有明显差异，2种抗原游离氨基酸含量在33％～40％之间，抗原蛋白含量均高于70％，血清效价在1：81920以上。结果提示，免疫抗原特异性较好。可用于单克隆抗体制备。

The immure, antigen and detecting antigen were synthesized with isothiocyanobenzyl -EDTA（ IEDTA ） , carrier prorein KLH or BSA and cadmium nitrate. Batb/C mice were immunized with the antigen to obtain antiserum. Two antigens were identiffed by three methods including ultraviolet speetrophotometry, trinitro- benzene -sulfonic acid （TNBS） and the autiseru m titer. The different ultraviolet absorbance were observed between synthesized antigen and carrier protein, the contents of free amino acid in antigen was from 33% to 40% , the titer of antiserum was above 1：81 920. These results indicated that the immune antigen with obvious specificity was an ideal candidate for monoclonal antibody preparation,