抗人甲状腺球蛋白单克隆抗体的制备及其对抗原决定簇特异性的研究

用杂交瘤技术建立三株分泌抗甲状腺球蛋白(TG)单克隆抗体的杂交瘤细胞株。所获三种单克隆抗体经兔抗小鼠IgG亚类、IgM及IgA血清鉴定表明,两种为IgG_1,一种为IgA;三种含单克隆抗体小鼠腹水的滴度(ELISA法)均为51200以上;以竞争性固相抗体结合试验测定它们对抗原决定簇的特异性差异,结果发现三种单克隆抗体的抗原结合部位是不同的,但对抗原的结合有一定的相互竞争抑制现象。

Preparation of human anti-thyrolobulin monoclonal antibodies and study on their specificity to antigenic epitopse

Three hybridoma cell lines secreting monoclonal antibodies (McAbS) against thyroglobulin (TG) were established by the fusion of NS-O mouse myeloma cells with the spleen cells of BALB/C mouse immunized with TG. Culture superna-tants were tested for specific antibodies with ELISA method. Cells from strongly positive hybrids were cloned twice. Ascitic fluids were prepared by inoculation of cloned hybridoma cells to BALB/C mouse imunized with TG. Culture super-natants were tested for specific antibodies with ELISA method. Cells from strongly positive hybrids were cloned twice. Ascitic fluids were prepared by inoculation of cloned hybridoma cells to BALB/C mice and purified by 33-50% ammonium sulfate precipitation. Three McAbs of the desired specificity were identified with rabbit antisera to mouse immunoglobnlin G (IgG1, IgG2a, Ig2b and IgG3) ,IgM and IgA,and the results showed that two of them were IgG1 ubtype,. one was IgA type. The titre of ascitic fluid for specific antibodies was above 51,200 by ELISA. The specificity to antigenic epitopes of these McAbs by competition solid phase antibody binding assay revealed that three McAbs might be bound to three different sites of the antigen, but there was competitive inhibition to some extent among themselves, probably caused by steric interference and these phenomena are expected to be proved by further study.