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从人心肌组织纯化酸性铁蛋白(AIF),免疫家兔产生的AIF血清用AIF偶联的亲和层析柱纯化,将抗AIF免疫球蛋白制成固相。125Ⅰ-抗AIF-H亚基单克隆抗体用氯胺-T法制备;采用顺序法建立AIF-H亚基因相免疫放射分析,数据用自动拟合样条函数处理,批内和批间CV分别为2.443%和10.160%,灵敏度为0.736μg/L,回收率为92.293%,可测范围为1.472-320μg/L,ED50为23.05μg/L。与AFP、CEA和乳铁蛋白(LF)无交叉反应,与铁蛋白有3.125%的交叉反应。健全性试验证明血清基质对本方法无干扰。40例男女血清AIF-H亚基含量分别为3.75±1.85和2.92±0.96μg/L(S±2S),10例肝癌患者血清AIF—H亚基含量为8.39±3.87μg/L(S±2S),显著高于正常参考值。本方法可为肿瘤,尤其是肝癌的基础和临床研究提供一种有效的手段。
Purified Acid Ferritin (AIF) from human cardiac muscle tissue, AIF sera from immunized rabbits were purified by AIF-coupled affinity chromatography to make anti-AIF immunoglobulin as a solid phase. The 125I-anti-AIF-H subunit monoclonal antibody was prepared by chloramine-T method. The AIF-H subunit immunofluorescence assay was established by sequential method. The data were processed by automatic fitting spline function. The intra- and inter- Of 2.443% and 10.160%. The sensitivity was 0.736μg / L, the recovery was 92.293%. The measurable range was 1.472-320μg / L, ED50 was 23.05μg / L. No cross-reactivity with AFP, CEA and lactoferrin (LF), 3.125% cross-reactivity with ferritin. Soundness tests demonstrate that serum matrices do not interfere with this method. The serum levels of AIF-H subunit in 40 cases were 3.75 ± 1.85 and 2.92 ± 0.96μg / L (S ± 2S), respectively. The serum levels of AIF-H subunit in 10 cases of HCC were 8.39 ± 3.87μg / L (S ± 2S), significantly higher than the normal reference value. The method can provide an effective method for the basic and clinical research of tumors, especially liver cancer.