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.
