抗慢性B淋巴细胞白血病患者血清IgM McAb的纯化及其酶结合物的制备

应用DEAE-22纤维素离子交换柱和正辛酸法从接种杂交瘤细胞BALB/C 鼠的腹水或腹腔肿瘤浸出液中纯化抗慢性淋巴细胞白血病(B-CLL)患者血清IgM 单克隆抗体(McAb),纯化物经巴比妥琼脂电泳,SDS-PAGE 鉴定为高纯度。ELISA 法证明具有高效价、高度特异性。采用过碘酸钠氧化结合法制备的辣根过氧化酶 HRPO-McAb 结合物的免疫活性、酶与蛋白质二者mol比、标记率均达到质量要求。

Purification of Monoclonal Antibodies against IgM from B-Chronic Lymphocytic Leukemia and Conjugation of HRPO to McAb

In this study, the anti-isotypic and anti-idiotypic antibodies (McAbs) against IgM of the patient with B chronic lymphocytic leukemia (B-CLL) were purified from the hy-bridoma ascites fluids or hybridoma supernatants by DEAE-22 cellulose ion-exchange chro-matography and n-Octoic acid precipitation method. The purification of the purified McAbs was evaluated by the immunoelectrophoresis and SDS-PAGE. High antibody activity and high specificity of purified McAbs were noted by use of ELISA. The conjugates of Horser-adish-peroxidase HRPO) to McAb were prepared by sodium metaperiodate method. The ex-perimental results showed that the ratio of peroxidase to McAb in conjugates and the effectiveness of conjugation were satisfactory. The conjugates of HRPO-McAb have a higher im-munoreactivity, as evidenced by double antibodies ELISA sandwich and indirect ELISA methods. The above findings indicate that the HRPO-McAb may be useful in characterizing McAb as well as quantitatively and qualitatively measuring immunoglobulin.