抗微囊藻毒素单克隆抗体的研制

Preparation of Monoclonal Antibody Against Microcystins

为研制抗微囊藻毒素(Microcystins,简称MC)的特异性单克隆抗体,用MC-KLH偶联物免疫6～8周龄雌性BalB/c小鼠,取脾细胞与小鼠骨髓瘤细胞Sp2/0融合,经过3～4次亚克隆建立稳定分泌抗MC的杂交瘤细胞株。将杂交瘤细胞注射入BalB/c小鼠腹腔,生产出抗MC的单克隆抗体。得到了抗微囊藻毒素单克隆抗体细胞株,命名为G5,抗体亚类为IgG2a,亲和力常数2.9×10-11mol/L,分子量150KD,加标回收率在81.5%～125.0%之间,与其他结构类似物的交叉反应率<1%。筛选出的抗微囊藻毒素单克隆抗体具有较好的特异性。

For development of monoelonal antibodies against Microcystin, a hybridoma cell line steadily secreting monoclonal antibodies against Microcystin , coded GS, was obtained by fusing murine Sp2/0 cells with spleen cells from mice immunized with MC-KLH conjugate and subcloning for 3 to 4 cycles. The ascites containing monoclonal antibodies against MC was gained via inoculation of the hybridoma cells into the abdominal cavity of BalB/c mice. The antibody subclass is IgG2a, affinity constant is 2.9×10^-11 mol/L, molecular weight is 150 kD, recovery is 81.5% - 125.0%, cross-reaction 〈 1%. The monoclonal antibodies thus prepared are highly sensitive and specific against microcystin.