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为开展弓形虫免疫诊断及抗弓形虫感染的相关研究,利用可溶性表达的弓形虫表面抗原SAG1截短型重组蛋白(trSAG1)研制单克隆抗体,筛选出能够稳定分泌高滴度抗体的杂交瘤细胞株,所分泌的单抗不仅适用于在虫体定位该蛋白,而且具有一定的小鼠体内保护作用。共筛选出4株(6H1、6G12、7B4和7A12)特异性好的单抗分泌株,经亚型测定,4株单抗重链均为IgG1,轻链均为κ链。抗体经纯化后测定效价为1∶8 000～1∶25 600;Western blot结果显示4株单抗能够同时识别重组表达及天然弓形虫SAG1抗原。筛选得到的2株抗SAG1单抗(6H1和7B4)可用于间接免疫荧光检测SAG1蛋白在虫体中的表达定位和分布。小鼠被动免疫保护试验表明,6H1和7B4两株单抗分别或联合使用可稍微延长被弓形虫感染的小鼠的存活时间。本试验研制的弓形虫SAG1单克隆抗体对将来研究弓形虫蛋白定位、弓形虫感染的诊断及防治具有一定的应用价值。
To carry out the immune diagnosis of Toxoplasma gondii and anti-Toxoplasma infections, the monoclonal antibody was developed by using soluble trichomonas surface antigen SAG1 trSAG1 to screen out the hybridoma cells that can stably secrete high titer antibody Strain, the secreted monoclonal antibody is not only suitable for positioning the protein in the worms, but also has some protective effect in mice. Four McAbs (6H1, 6G12, 7B4 and 7A12) were screened for their specific secreted McAbs. Based on the subtypes, the four McAbs were all IgG1 in the heavy chain and the κ chain in the light chain. The antibody titer was 1: 8000 ~ 1:25 600 after purification. Western blot results showed that the four McAbs could recognize both recombinant expression and Toxoplasma gondii SAG1 antigen simultaneously. Two anti-SAG1 monoclonal antibodies (6H1 and 7B4) were screened and used for the indirect immunofluorescence assay to detect the expression and distribution of SAG1 protein in the worms. Passive immune protection test in mice showed that the two monoclonal antibodies 6H1 and 7B4 could prolong the survival time of mice infected with Toxoplasma gondii, respectively or in combination. The test developed Toxoplasma gondii SAG1 monoclonal antibody for the future study of Toxoplasma protein localization, Toxoplasma gondii infection diagnosis and prevention and treatment has a certain value.