抗麻痹性贝毒素GTX2,3单克隆抗体的制备及特性分析

制备抗麻痹性贝毒GTX2,3单克隆抗体。利用醛化法将GTX2,3与载体牛血清白蛋白(BSA)偶联,制备完全抗原。免疫小鼠,取小鼠脾细胞与Sp2/0细胞融合。GTX2,3与钥孔血蓝蛋白(KLH)偶联作为检测抗原,用间接ELISA法筛选阳性克隆株。将筛选的阳性细胞株制备腹水。获得三株稳定分泌抗GTX2,3单克隆抗体的杂交瘤细胞株F4、F10、G9。间接ELISA法检测F10细胞株腹水抗体效价为1.4×10^-5。半抗原GTX2,3与载体蛋白偶联后,作为免疫原,可制备高滴度的抗GTX2,3抗血清和单克隆抗体。该抗体对于藻毒素具有高特异性和高亲和力,可用于污染海产品的麻痹性贝毒的检测。

Preparation and characterization of monoclonal antibodes against Paralytic Shellfish Poisoning Toxins GTX2, 3

To obtain hybridoma cell line which can secrete anti-gonyautoxin 2,3(GTX2,3) monoclonal antibodies(mAb), the immunogen(GTX2,3-BSA) and solid phase immunogen(GTX2,3-KLH) were prepared by the method of aldehyde conjugation, in which GTX 2,3 was conjugated with BSA and KLH respectively. The spleen cells from BALB/c mice immunized with GTX2,3-BSA were fused with murine Sp2/0 myeloma cells in PEG4000. The specific hybridoma cells and the ascites were analyzed by ELISA. Three specific hybridoma cell lines which can secrete anti-GTX 2,3 mAb were obtained. The titer of the mAb in the ascites was about 1.4×10^-5. The specific anti-GTX2,3 mAb can be obtained from hybridoma successfully, which can used to detect the PSP from seafood.