绒毛膜促性腺激素受体单克隆抗体的制备及其特性的初步研究

preparation of Monoclonal Antibodies Against Chorionic Gonadotropin Receptor and Study of Its Characteristics

以纯化嗜麦芽假单胞菌的绒毛膜促性腺激素(CG)受体为抗原,融合得到4株(ED490,DG390,AB890,GE590)对细菌CG受体特异的单克隆抗体杂交瘤细胞,其腹水和培养上清中的抗体滴度分别为10^(-2)～10^(-6)和1～10^(-2);亚类鉴定显示:单抗GE590为IgGl,单抗ED490、DG390和AB890为IgG2b。研究发现,ED490、AB890和DG390与细菌CG受体作用后,^(125)I-HCG仍能结合该受体,揭示其作用于受体不同位点。GE590与CG受体结合实验发现,随着单抗浓度不断增加,^(125)I-HCG与受体结合逐渐减少,推测该单抗与^(125)I-HCG结合受体同一位点。

We reported the productionof monoclonal antibodies (McAbs) againstchorionic gonadotropin hormone (CG) re-ceptor by fusing spleen cells of BALB/cmice which had been immunized by puri-fied bacteria (Pseudomonas maltophilia)CG receptor with mouse myeloma lineSP2/0. Four hybridoma cel1 lines secretingCG receptor McAbs were obtained (ED490DG390, AB890 and GE590). The titers ofspecific antibodies of both mice ascites andculture supernatant were 10^(-2)-10^(-5) and1-10^(-2) respectively, by solid phase ELISA.Double-Immunodiffusion test showed thatthe McAb GE590 was IgG_1,and the McAbsED490, DG390 and AB890 were IgG2b Immu-noprecipitation indicated that ^(125)I-HCGcould bind the HCG receptor which had re-acted with McAbs ED490, AB890 and DG390,suggesting that they may recognize the re-ceptor with different antigenic datermi-nant. Interaction of McAb GE590 with thereceptor showed that the increased concen-tration of GE590 was in inverse propor-tion to the amount of ^(125)I-HCG binding re-ceptor, indicating that both the McAb and^(125)I-HCG could recognize a common site ofreceptor and that increased concentrationof McAb GE590 may induce some changein conformation and structure of the re-ceptor. Our study suggested that theseMcAbs may be used for studying structureof CG receptor.