论文部分内容阅读
用本室纯化的人前列腺特异抗原免疫ＢＡＬＢ／ｃ小鼠，经淋巴细胞杂交瘤技术，建立了４株体外稳定分泌抗人前列腺特异抗原的杂交瘤细胞株，分别命名为ＱＷ２、ＱＷ４、ＱＷ７和ＱＷ８。将杂交瘤细胞注入同系小鼠腹腔制备腹水，用ＥＬＩＳＡ方法测得腹水抗体效价为１０６，其中ＱＷ８的抗体滴度最高。ＱＷ８腹水经５０％硫酸铵沉淀后包被酶标板，建成双抗体夹心ＥＬＩＳＡ，标准曲线线性良好，灵敏度为１．０μｇ／Ｌ血清稀释曲线，回收率及批内、批间变异系数测定表明，该方法的准确性和精密度均符合质量控制要求。检测８１名５０岁以下正常男性血清，ＰＳＡ均值为１．０５±０．７３μｇ／Ｌ，１０例临床确诊为前列腺癌患者的血清ＰＡＳ均值为３６．５±２．３μｇ／Ｌ明显高于正常值。
BALB / c mice were immunized with purified human prostate specific antigen and four hybridoma cell lines secreting anti-human prostate specific antigen were established by lymphocyte hybridoma technology and named QW2, QW4, QW7 and QW8. The hybridoma cells were injected into the peritoneal cavity of homologous mice to prepare ascites. The antibody titer of ascites was 106 by ELISA, of which the highest antibody titers of QW8 were obtained. The ascites of QW8 was precipitated with 50% ammonium sulfate and then coated with ELISA plate. The double antibody sandwich ELISA was established. The calibration curve was linear and the sensitivity was 1.0 μg / L. The recovery curve and the variation coefficient of intra- and inter- The accuracy and precision of the method are in line with the quality control requirements. Serum PSA of 81 men under 50 years of age was detected with a mean of 1.05 ± 0.73 μg / L. The mean serum PAS value of 10 clinically diagnosed prostate cancer patients was 36.5 ± 2.3 μg / L, which was significantly higher than the normal value .