抗人IgD单克隆抗体的制备及其特性鉴定

本文采用Ultrogel AcA34凝胶过滤,DEAE离子交换及亲和层析法,从骨髓瘤病人血清中分离高纯度的IgG,以此为抗原免疫小鼠,利用常规杂交瘤技术制备了3株分泌抗人IgD单克隆抗体的杂交瘤细胞株,分别命名为ZD2、ZD61和ZD94,此3株杂交瘤细胞分泌的抗体与IgD有特异性反应,而与人其它类别的免疫球蛋白重、轻链无交叉反应,均属IgG1亚类。免疫转印结果显示,均能识别分子量为67kD的IgD重链条带。用ELISA法测定3株抗体分别识别两种不同的抗原决定簇。

Production and Characterization of Monoclonal Antibodies against Human IgD

In the present study, human IgD was purified from the serum of a patient with myeloma through gel filtration on Ultrogel AcA34, DEAE ion-exchange chromat-ography and immunoaffinity chromatography. The prepared IgD was used as antigen and conventional hybridoma technique was employed to raise monoclonal antibodies (McAbs) against human IgD, Three cell lines capable of steady production of McAbs. were established and they were designated ZD2, ZD61 and ZD94, respectively. Determination of immunoglobulin class and subclass showed that these hybridoma cultures synthesized mouse IgGl isotype, which reacted only with human IgD and did not react with the light and heavy chains of other human immunoglobulins. The immunoblotting results indicated that these McAbs recognized one human IgD heavy chain band with a molecular weight of 67 kD. Two different epitopes of human IgD were defined by these 3 McAbs using ELISA.