Stamenova M, Tzvetkova V, Kehayov I, Kyurkchiev S
Department of Molecular Immunology, Bulgarian Academy of Sciences, Sofia, Bulgaria.
Int J Biochem Cell Biol. 1995 Nov;27(11):1117-22. doi: 10.1016/1357-2725(95)00099-b.
Monoclonal antibody designated 1B10 (Mab 1B10) has been shown to be highly specific for the beta-chain of human chorionic gonadotrophin (HCG). We used this antibody to investigate its paratope using anti-idiotypic antibodies. Purified Mab 1B10 has been used to immunize syngeneic BALB/c mice to produce anti-idiotypic monoclonal antibodies. An enzyme immunoassay (ELISA) on Mab 1B10 coated plate was employed to screen the supernatants of growing hybridomas. The specificity of each antibody selected was assessed using an inhibition ELISA and immunoblotting. Monoclonal antibodies belonging to two categories were selected. (a) Those (designated Mab 4F8 and Mab 7G9) recognizing epitopes of the Ig molecule located in/or near the antigen-binding site of Mab 1B10. In ELISA these antibodies were shown to inhibit in a dose-dependent manner, the reaction of Mab 1B10 with its specific antigen; (b) those (Mab 2B8, Mab 3B8) reacting with epitopes located outside of the antigen binding site of the antiHCG antibody molecule and did not influence the reactions of Mab 1B10 and its antigen. Following immunization of syngeneic BALB/c mice monoclonal antibodies (Mab 4F8, Mab 7G9) were produced which recognized epitopes located on the variable region of Mab 1B10 since they did not react with other murine monoclonal antibodies of the same isotype. These antibodies inhibited the binding of Mab 1B10 to its corresponding epitope on the molecule of HCG and they can be defined as syngeneic anti-idiotypic antibodies.
名为1B10的单克隆抗体（Mab 1B10）已被证明对人绒毛膜促性腺激素（HCG）的β链具有高度特异性。我们使用该抗体，通过抗独特型抗体来研究其互补决定区。纯化的Mab 1B10已用于免疫同基因BALB/c小鼠，以产生抗独特型单克隆抗体。采用在包被有Mab 1B10的平板上进行酶免疫测定（ELISA）来筛选生长中的杂交瘤细胞的上清液。使用抑制ELISA和免疫印迹法评估所选每种抗体的特异性。选择了两类单克隆抗体。（a）那些（命名为Mab 4F8和Mab 7G9）识别位于Mab 1B10抗原结合位点内/附近的Ig分子表位的抗体。在ELISA中，这些抗体呈剂量依赖性地抑制Mab 1B10与其特异性抗原的反应；（b）那些（Mab 2B8、Mab 3B8）与抗HCG抗体分子抗原结合位点外的表位发生反应且不影响Mab 1B10与其抗原反应的抗体。在免疫同基因BALB/c小鼠后，产生了识别位于Mab 1B10可变区表位的单克隆抗体（Mab 4F8、Mab 7G9），因为它们不与相同亚型的其他鼠单克隆抗体发生反应。这些抗体抑制Mab 1B10与其在HCG分子上相应表位的结合，可被定义为同基因抗独特型抗体。
