Lou J, Guo Z, Meng Z
Institute of Nutrition and Food Hygiene, Chinese Academy of Preventive Medicine, Beijing.
Zhonghua Yu Fang Yi Xue Za Zhi. 1995 Mar;29(2):92-5.
Compound antigen, bovine serum albumin-sterigmatocystin (BSA-ST) or hemocyanin sterigmatocystin (H-ST) was prepared by converting ST to its hemiacetal group in ST and taking ST conjugated to BSA or H. Balb/c mice were immunized with BSA-ST, and then their spleen cells were isolated and fused with SP2/0 murine myeloma cells. A hybridoma cell strain that could stably secrete monoclonal antibody against ST (McAb-ST) was selected, and both of their ascites and serum were rich in McAb-ST induced by cells of this strain. McAb-ST was identified as Ig-Gi with a molecular weight of 168,000 kd (53,000 for heavy chain and 31,000 for light one), and with an affinity constant of H-ST reaction of 1.29 x 10(9) mol/L measured by indirect competitive enzyme-linked immunosorbent assay (IC-ELISA). Relative cross reaction of McAb-ST with ST analogues was less than 2%. The minimal 0.5 pg/25 microliters of ST can be detected with IC-ELISA using McAb-ST and ST simultaneously, with a linear range of 0.1-10 ng/ml.
通过将柄曲霉素（ST）转化为其半缩醛基团，并使ST与牛血清白蛋白（BSA）或血蓝蛋白（H）偶联，制备了复合抗原，即牛血清白蛋白 - 柄曲霉素（BSA - ST）或血蓝蛋白 - 柄曲霉素（H - ST）。用BSA - ST免疫Balb / c小鼠，然后分离其脾细胞并与SP2 / 0鼠骨髓瘤细胞融合。筛选出能够稳定分泌抗ST单克隆抗体（McAb - ST）的杂交瘤细胞株，该细胞株诱导产生的腹水和血清中均富含McAb - ST。McAb - ST被鉴定为Ig - Gi，分子量为168,000 kd（重链为53,000，轻链为31,000），通过间接竞争酶联免疫吸附测定（IC - ELISA）测得其与H - ST反应的亲和常数为1.29×10⁹ mol/L。McAb - ST与ST类似物的相对交叉反应小于2%。使用McAb - ST和ST同时进行IC - ELISA时，最低可检测到0.5 pg/25微升的ST，线性范围为0.1 - 10 ng/ml。
