论文部分内容阅读
目的:制备人绒毛膜促性腺激素(β-HCG)单克隆抗体,建立人β-HCG双抗体夹心CLIA检测方法。方法:用人β-HCG抗原免疫小鼠,通过细胞融合、筛选后得到杂交瘤细胞株,然后将细胞株扩大培养并纯化上清液获得抗体,测定抗体亲和力、特异性及表位,最后建立双抗体夹心CLIA方法。结果:获得4株抗人β-HCG的杂交瘤细胞株(β-1-1、β-2-1、β-3-1、β-4-1)。用β-1-1和β-2-1建立的双抗体夹心法的检测范围为0.5 mIU/mL-800 mIU/mL,灵敏度0.23 mIU/mL,检测结果的相对偏差均在±10%内,回收率在90%以上。结论:本研究最终成功制备了抗人β-HCG mAb,建立了定量检测人β-HCG的双抗体夹心CLIA方法,为β-HCG检测及疾病的诊断奠定基础。
OBJECTIVE: To prepare a human chorionic gonadotropin (β-HCG) monoclonal antibody and to establish a human β-HCG double antibody sandwich CLIA assay. Methods: Mice were immunized with human β-HCG antigen, and the hybridoma cell lines were screened by cell fusion. Then the cell strains were expanded and purified to obtain antibodies. The affinity, specificity and epitope of the antibodies were determined. Finally, bis Antibody sandwich CLIA method. Results: Four anti-human β-HCG hybridoma cell lines (β-1-1, β-2-1, β-3-1, β-4-1) were obtained. The detection range of the double antibody sandwich method with β-1-1 and β-2-1 was 0.5 mIU / mL-800 mIU / mL and the sensitivity was 0.23 mIU / mL. The relative deviation of the detection results was within ± 10% The recovery rate is over 90%. Conclusion: The anti-human β-HCG mAb was successfully prepared in this study. A double antibody sandwich CLIA method for quantitative determination of human β-HCG was established, which laid the foundation for the detection of β-HCG and the diagnosis of the disease.