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从人心肌组织中提纯了酸性铁蛋白,免疫家兔产生抗血清,氯胺-T法制备了~(125)Ⅰ-酸性铁蛋白和~(125)Ⅰ-抗酸性铁蛋白H亚基单抗。分别采用平衡法和顺序饱和法建立了酸性铁蛋白放射免疫分析和酸性铁蛋白H亚基免疫放射测量分析。它们的批内CV分别为1.65％和2.44％,批间CV为9.71％和10.16％,回收率为102％和92.29％,ED_(50)为27.50μg/L和23.05μg/L。前者的可测范围为7.0～369.6μg/L,后者的灵敏度为0.736μg/L。两方法与甲胎蛋白、癌胚抗原和乳铁蛋白均无交叉反应,与铁蛋白的交叉反应分别为1.74％和3.13％。分析各年龄组正常人血清酸性铁蛋白含量发现其受性别和年龄影响;酸性铁蛋白分析肝癌的敏感度为73.05％,基本不受非癌性肝病的干扰;八种恶性肿瘤诊断试验评价结果显示,酸性铁蛋白含量对肝癌的诊断价值最高。酸性铁蛋白H亚基分析表明:在肝癌患者血清中的含量显著高于正常参考值。建议将上述两种方法与甲胎蛋白分析进行平行试验,提高肝癌的诊断水平。
Purification of acidic ferritin from human myocardial tissue, immunization of rabbits to produce antiserum, preparation of 125 I-acid ferritin and 125 I-anti-acidic ferritin H subunit mAb by chloramine-T method . The radio-immunoassay of acid ferritin and the immunoradiometric analysis of acid ferritin H subunit were established using the equilibrium method and sequential saturation method, respectively. Their intra-assay CVs were 1.65% and 2.44%, inter-assay CVs were 9.71% and 10.16%, and the recoveries were 102% and 92.29%, respectively. ED_(50) was 27.50 μg/L and 23.05 μg/L. The former has a measurable range of 7.0-369.6 μg/L and the latter has a sensitivity of 0.736 μg/L. The two methods did not cross-react with alpha fetoprotein, carcinoembryonic antigen, and lactoferrin, and the cross-reactivity with ferritin was 1.74% and 3.13%, respectively. Analysis of serum ferritin levels in normal people of each age group was found to be affected by gender and age. The sensitivity of acid ferritin analysis to hepatocellular carcinoma was 73.05%, which was basically unaffected by non-cancerous liver disease. Eight malignancy diagnosis test results showed that The content of acid ferritin has the highest diagnostic value for liver cancer. The analysis of the acidic ferritin H subunit showed that the content of serum in liver cancer patients was significantly higher than the normal reference value. It is recommended that the above two methods be performed in parallel with alpha-fetoprotein analysis to improve the diagnostic level of liver cancer.