孕马血清促性腺激素单克隆抗体的研制

STUDIES OF MONOCLONAL ANTIBODIESAGAINST PREGNANT MABE SEBUM GONADOTBOPIN

用高纯度孕马血清促性腺激素免疫ＢＡＬＢ／ｃ小鼠，加完全佐剂进行基础免疫，第７日脾脏免疫，第１０日取血测滴度，为阳性则取出脾脏。将脾细胞与Ｓｐ２／０细胞融合，培养并用琼脂单扩法筛选抗体生成孔，ＥＬＩＳＡ复测，得孕马血清促性腺激素抗体阳性杂交瘤细胞株Ｄ＿６Ｃ＿（１０）。经降植烷予处理的ＢＡＬＢ／ｃ鼠腹腔注射Ｄ＿６Ｃ＿（１０），腹水单抗滴度１０￣６。瘤细胞ＤＮＡ含量测定证实为脾和骨髓瘤细胞的杂交瘤细胞。单抗亚型分析为ＩｇＭ型。建立了间接酶免疫法快速测定孕马血清促性腺激素单克隆抗体的滴度。

BALB/C mice were immunized using highly purified PMSG with complete Fruend'sadjvant as basic immunization. A week later spleen immunization was performed. Afterthree days, the mice blood samples were tested for antibody titer. If it was positive,mice were killed and the cell fusion between spleen cells and Sp_(2/0) were made and aPMSG McAb hybridoma cell line produced. The hybridoma supernatants were detected,the positive ones were subcloned and further cultured. Ascitic fluids were induced afterinoculating positive hybridoma cells D_6C_(10) into abdominal cavity of BALB/c mice whichpretreated with Pristane. The titer of ascitic fluids was more than 10￣6. By DNA analysis.hybridoma cells D_6C_(10) producing PMSG McAb were proved to be hybrid cells. Screeningthe subclass of McAb, the precipitation line appeared to be IgM. A matrix cross mat-ching experiment was conducted to select the titer of PMSG McAbs. An indirect enzyme-linked immunosorbent assay method for measurment of PMSG McAb titer was establi-shed .