抗重金属铜离子单克隆抗体的制备及其免疫学特性

Preparation and Immunology Character of Monoclonal Antibody of Heavy Metal Ion Copper

为制备抗Cu^(2+)的高效价、敏感、特异的单克隆抗体,采用已合成的铜离子人工抗原Cu^(2+)-ITCBEBSA免疫Balb/C小鼠,通过间接ELISA和阻断ELISA筛选备用小鼠,用细胞融合技术制备抗Cu^(2+)的单克隆抗体。结果筛选到4株特异性强、灵敏度高的杂交瘤细胞,其中杂交瘤细胞株2B8培养上清中的抗体效价采用间接ELISA检测为1∶(1.28×10~3),用其制备的腹水中抗体效价为1∶(5.12×10~5),该单克隆抗体为IgG1/κ型,对Cu^(2+)-EDTA的半数抑制质量浓度(IC_(50))为29.78μg/L,与Zn^(2+)-EDTA的交叉反应率(CR)为26.0%,与其他金属螯合物没有交叉反应,表明获得特异性较好的抗Cu^(2+)的单克隆抗体,可用于铜离子的免疫学检测。

To produce efficient, sensitive and specific monoclonal antibody, the experimental mouse was selected from the composited Cu^2＋-ITCBE-BSA to immunize Balb/C mouse by the means of indirect ELISA and blocking ELISA, and monoclonal antibody was prepared by cell fusion technology. The results showed that four hybridoma cell lines characterized as powerful specificity and high sensi- tivity were obtained. The indirect ELISA titers of 2B8 were 1： （1.28 ×10^5） in supernatant and 1∶（5.12×10^5） in ascites. The isotype of the 2B8 mAb was IgG1/k. The Cu2＋-EDTA-mAb showed that the IC50 of mAb to Cu^2＋-EDTA was 29.78 μg/L, 26.0% cross-reactivity to Zn^2＋-EDTA and little or no cross-reactivity with other compounds. The prepared monoclonal antibody of Cu^2＋-EDTA can be used for immunoassay of Cu^2＋.