猕猴黄体生成素放射免疫分析方法的建立

Development of a method for RIA of MLH

目的 :在生殖医学的动物实验中 ,需要检测猕猴血清中黄体生成素 (MLH)的含量 ,为此建立了猕猴血清黄体生成素 (MLH)的放射免疫分析方法。方法 :用氯胺T法制备12 5I-MLH ,兔抗MLH为第一抗体 ,羊抗兔IgG为分离剂 ,采用液相竞争法建立放射免疫分析方法。结果 :标记抗原比放射性为 6 7μg/mCi,抗体亲和常数为Ka=3 5 5× 10 -9mol/L ,标准曲线形态良好 ,r=0 991,批内误差CV =3 49% ,批间误差CV =4 6 5 % ,最小可测浓度为 0 42 μg/ml,30例正常猕猴血清检测结果为 1 17± 0 6 75 μg/ml。 结论 :MLHRIA的建立对避孕药和性生理药品的动物监测及医学研究均有重要意义。

Objective To develop a method of RIA for MLH to measure the serum level of LH in Rhesus Monkey for animal expeiment in reproductive medicine. Methods 125 I-MLH was prepared with chloramine T-Iodogen method. Rabbit anti-MLH was used as first antibody and sheep anti-rabbit IgG was used as separating agent. RIA was parformed with liquid phase competitive radioassay. Results The specific radioactivity of labelled antigen was 67μg/mCi; antibody affinity constant was Ka=3.44×10 -9 mol/L the shape of the standard curve was good. r=0.991 , intra-assay error CV=3.49%, inter-assay error CV=4.65%. The mininal detectable concentration was 0.42μg/ml. Mean value of 30 normal Rhesus Monkey serum specimens was 1.17±1.30μg/ml. Conclusion The developed method was simple reliable and sensitive. It would be of use in study of contraceptive and sex-physiology drugs.