摘要
采用淋巴细胞杂交瘤技术,获得了5株稳定分泌抗促肾上腺皮质激素释放因子(CRF)的McAb杂交瘤细胞株。5株单克隆抗体均为IgG2,且均识别CRF羧基端。ELISA交叉试验结果表明,该McAb不与ACTH、FSH、NPY、LH反应。5株杂支瘤腹水效价在10-4~10-5,亲和常数在5×107mol/L-1~1×109mol/L-1。
A new immunization method for preparation of monoclonal antibody(McAb) against corticortropin-releasing factor(CRF),that is directly emulsifing synthetic CRF with completed Frenud's adjuvant,and then injecting BALB/c mice intraperitoneally every 2 weeks with 1 μg og CRF.Using lymphocyte hybridoma technique,5 lines of hybridomas stably secreting McAb against CRF were obtained by the screening hybridoma culture supernatant in Enzyme-Linked Immunosorbent assay (ELISA) with CRF on Solid phase. All of McAb were IgG2 class and combined with carboxyl terminal of CRF.Cross-reactivities with ACTH, FSH, NPY and LH were invisible using the ELISA for CRF.Titration of hybridomas culture supernatant were between 1:200 to 1:400, and titration of ascitic fluid of hybridomas were between 1:10 000 to 1:100 000.Their affinities constants were 1 X 108mol/L-1,5X 107 mol/L-1,1 X 108 mol/L-1, 1 X 109 mol/L-1 and 1X 109 mol/L-1 respectively.