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[抗柄曲霉素单克隆抗体的制备与鉴定]

[Preparation and identification of monoclonal antibody against sterigmatocystin].

作者信息

Lou J, Guo Z, Meng Z

机构信息

Institute of Nutrition and Food Hygiene, Chinese Academy of Preventive Medicine, Beijing.

出版信息

Zhonghua Yu Fang Yi Xue Za Zhi. 1995 Mar;29(2):92-5.

PMID:7796690
Abstract

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.

摘要

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。

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