人精子蛋白17放射免疫分析方法的建立

目的:建立人精子蛋白17(Sp17)的放射免疫分析方法。方法:用重组人Sp17(rhSp17)免疫动物制备高质量的抗血清,用氯胺T法制备125I-rhSp17,建立了Sp17放射免疫分析方法,并测定了正常人(n=59)及35例患者(卵巢癌20例,子宫癌14例,宫颈癌1例)血清Sp17值。结果:测定范围3.3~800μg/L;灵敏度为2.0μg/L;批内CV为7.5%~9.8%,批间CV为8.2%~13.2%。正常人血清Sp17值为(15.60±7.66)μg/L。以Sp17>31.92μg/L为阳性,35例女性肿瘤患者血清标本15例阳性。结论:Sp17的放射免疫分析是一种简便、灵敏、特异的分析方法,可满足临床和科研的需要;Sp17的测定对研究该蛋白的功能、天然分布和异常表达有重要意义。

Development of Radioimmunoassay for Human Sperm Protein 17

OBJECTIVE: To develop a method of radioimmunoassay for human sperm protein 17(Sp17) determination. METHODS: Anti-recombinant human Sp17 antibody was prepared, the labeling of 125I-rhSp17 performed by chloramine T method, and radioimmunoassay of Sp17 developed. Results: The assay range was 3.3 to approximately 800 microg/L, the sensitivity was 2.0 microg/L, and the intra- and inter-assay coefficients of variation (CV) were 7.5% to approximately 9.8% and 8.2% to approximately 13.2%, respectively. The serum Sp17 level in normal subjects was (15.60 +/- 7.66) microg/L (n = 59). CONCLUSION: This radioimmunoassay of Sp17 fulfills the reasonable requirements of clinical routine and scientific studies in terms of specificity, sensitivity and practicability. Measurement of Sp17 concentration is useful for assessing its native distribution and aberrant expression.