脂肪酸结合蛋白放射免疫分析方法的建立及初步应用

The Establishment and Primary Application of Specific Fatty Acid Binding Protein Radioimmunoassay

用人工重组的人脂肪酸结合蛋白 (FABP)多次免疫大耳白兔 ,获取高效价的FABP抗体。用氯胺T法制备12 5I-FABP ,再经SephadexG - 2 5纯化 ;抗原抗体反应采用平衡一步法 ,4℃培养 2 4h后 ,经PR试剂分离结合和游离的标记抗原。该法测定范围 5 - 4 0 5ng/mL ,最低检出量为 9ng/mL ,批内和批间CV分别小于 6 .4 %和8.5 %。用该法测得人血清FABP含量 :健康人 (4 5名 )为 15 .6 8± 2 .91ng/mL ;重症监护病人 (4 6例 )为 72 .0 8±32 .6 4ng/mL ;心脏病患者 (73例 )为 78.95± 2 4 .83ng/mL ,明显升高。结果表明 ,本法稳定 ,简便特异 ,其灵敏度适于检出人血清FABP水平的变化。

A highly specific and highly sensitive radioimmunoassay method for fatty acid binding protein (FABP) in human serum was developed.The highly effective antibody againist FABP was obtained by immunizing rabbits with human recombinaed FABP.The FABP was labeled with 125 I by chloramines-T methods and purified by the Sephadex-G25 column.Te reaction between antigen and antibody was carried out by one step balance method and incubated in 4℃ for 24 hours, then binding and free antigen were separated by PR reagent.The detection range of this method is from 5 to 405ng/mL;the lowest detection level is 9ng/mL.CV′s within batche and between batch were less than 6.4% and 8.5% respectively.The serum FABP concentration of healthy persons is 15.68±2.91ng/mL(n=45),that of ICU patients is 72.08± 32.64 ng/mL(n=46) and that of cardiopathy patients is 78.95±24.83ng/mL(n=73).It suggest that this method is stable, simple, specific and highly sensitive.It is suitable for testing FABP in human serum.