前列腺特异抗原的酶联免疫分析

Enzyme-linked Immunosorbent Assay for Prostate Specific Antigen

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。

Four hybridoma cell strains secreting monoclonal antibodies(McAbs QW1,QW4,QW7 and QW8) were obtained by immunizing mice with human PSA prepared in our laboratory.Hybridoma cell strains injected into the abdominal cavity of mice produced four McAbs with a titer of 10￣6,that of QW8 being the highest. Two-site sandwich enzyme-linked immunosorbent assay(ELISA) has been developed by the binding of the McAb coated on a microplate with biotinylated PcAb. Its sensitivity was 1. 0μg/L. The standard curve,accuracy and precision of the assay were found to comform to the quality control demands through the evaluation of serum dilution curve ,recovery rate and coefficients of variation. The mean of serum PSA in 10 patients with carcinoma of the prostate was 36. 5±2. 3μg/L ,significantly higher than that of the normal males.