胰岛素样生长因子-1的放射免疫测定方法的建立

Development of a radioimmunoassay for insulinlike growth factor1

胰岛素样生长因子－１（ＩＧＦ－１）分别与两种载体蛋白——牛血清白蛋白（ＢＳＡ）和卵清蛋白（ＯＡ）用戊二醛连接，经紫外分光光度仪扫描确定偶联成功。以ＩＧＦ－１－ＢＳＡ作为免疫原，皮内多点注射免疫青紫兰兔，制备ＩＧＦ－１抗体，经ＥＬＩＳＡ法测定，血清效价为１∶２０００，放免工作浓度为１∶１５０００。采用氯胺－Ｔ标记ＩＧＦ－１，用酸醇抽提法处理样品，去除ＩＧＦ－１结合蛋白。所建立的ＩＧＦ－１放免标准曲线，其最小检测量为０．０６ｎｇ／ｍＬ，批内误差为４．８％，批间误差为８．２％（ｎ＝１０）。

rhIGF1 and BSA(OA) was conjugated with glutaraldehyde.The prepared complete antigen BSAIGF1 and OAIGF1 were confirmed by ultraviolet spectrometry techniques.Two Chinchilla rabbits were immunized by multiple site subcutaneous and intradermal injection of BSAIGF1.The antiserum exhibited an effective titer of 12 000 by ELISA and 115 000 by RIA.IGF1 was iodinated using chloramineT.Acidethanol extraction of samples was to separate and precipate IGF1 binding protein for assay.The standard curve was established using DNArecombinant human IGF1 standard with a calculated sensitivity of 006 ng/mL,where the intraassay variance and interassay variance were 48% and 82%,respectively( n =10).