特异性白介素1受体拮抗蛋白放射免疫分析的建立及初步应用

The Establish and Primary Application of Specific Interleukin-1 Receptor Antagonist Protein Radioimmunoassay

用人工重组的白介素 1受体拮抗蛋白 (I 1ra)免疫新西兰兔 ,获取高效价特异的IL 1ra抗体。用氯氨T氧化法制备12 5I标记IL 1ra ,经SephadexG 2 5纯化 ,竞争性抗原抗体反应采用非平衡法 ,标准品和待测样品同抗体反应 37℃ 6小时后加入12 5I 标记抗原继续反应 4℃ 2 4小时。经PR试剂分离结合和分离的标记抗原。该法标准曲线范围 3～ 96ng ml,最低检出量为 ( 9 0± 1 4ng ml,n =89) ,类风湿因子阳性的病人血清含量为 ( 39 0 6± 15 2 3ng ml,n =14)。肠缺血再灌损伤后 6小时大鼠血清中IL 1ra较伤前自身对照明显增加 (P <0 0 0 1,n =82 )。肺灌洗液中损伤前后没有明显差异。该法特异、灵敏、简便 ,可用于人血清和大鼠血清及肺灌注液中IL

The New Zealand rabbits were immunolized with rhIL\|Ira and obtained high titer specific IL\|1ra antibody.The 125 I IL 1ra was prepared by chlormines T methods and purified by the Sephadex\|G25 chromatograph column.The competing reaction between antigen and antibody was carried out by unbalanced method and standards and samples mixed with antibody firstly incubated for 6v hours at 37℃,following added 125 I labbled IL 1ra for 24 hours at 4℃,then separated bond and free antigen by PR reagent.The detection range of this method was about 3～96ng/ml,the lowest detection level was 1ng/ml,error within batches and between batches was less than 5\^3% and 10%.The serum IL 1ra concentration in normal peoples was 9\^01±1\^4ng/ml(n=89),Ra factor positive patients serum IL 1ra was 39\^06±15\^23ng/ml,(n=14).IL 1ra level was increased significantly in rat serum after 6 hours of intestinal ischemia reperfussion injury compared with self\|control before injury (4\^04±0\^71ng/ml vs 5\^53±1\^33 P<0\^001, n=82).IL 1ra level in lung lavage was without obvious changes.This method was specific,sensitive,and simple,which could be used in human and rat serum or lung lavage for IL 1ra analysis.