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制备配对胃蛋白酶原Ⅱ(PGⅡ)单抗,建立人血清中胃蛋白酶原Ⅱ夹心ELISA检测方法。用胃蛋白酶原Ⅱ免疫Balb/c小鼠,制备免疫脾细胞,与SP2/0融合,用HAT培养基进行筛选培养,间接ELISA检测阳性克隆,对阳性孔进行多次单克隆化,选出效价高、分泌性能稳定的杂交瘤细胞,制备腹水并进行纯化。进行单抗配对,建立胃蛋白酶原Ⅱ双抗体夹心检测方法。获得1D8、1C6、2H11、2E3等4株杂交瘤,经配对试验,确定1D8、2H11可作为夹心ELISA检测PGⅡ的单抗。成功制备出配对单抗,初步建立了PGⅡ双抗体夹心ELISA方法。
Preparation of paired pepsinogen Ⅱ (PG Ⅱ) monoclonal antibody, the establishment of human serum pepsinogen Ⅱ sandwich ELISA detection method. Balb / c mice were immunized with pepsinogen II, immunized spleen cells were prepared, fused with SP2 / 0, screened and cultured in HAT medium, positive clones were detected by indirect ELISA, multiple positive clones were selected, and selected for efficacy High price, secretion of stable hybridoma cells, preparation and purification of ascites. Paired mAb, the establishment of pepsinogen Ⅱ double antibody sandwich detection method. Obtained 1D8, 1C6, 2H11, 2E3 other 4 hybridomas, the paired test to determine 1D8, 2H11 can be used as a sandwich ELISA detection of PG II monoclonal antibody. Paired monoclonal antibodies were successfully prepared, and a sandwich enzyme-linked immunosorbent assay (ELISA) was established.