摘要
目的:建立高特异性和高灵敏度的人胰岛素样生长因子Ⅱ(IGF-Ⅱ)放射免疫分析方法,用于测定人血清以及培养液中IGF-Ⅱ水平。方法:用人工合成的IGF-Ⅱ多肽连接甲状腺球蛋白,多次免疫兔子获取高效价、高特异的人IGF-Ⅱ抗体。用氯氨T法制备125I标记的IGF-Ⅱ,经交联葡聚糖凝胶(Sephadex G-25)纯化。抗原抗体反应采用平衡一步法,4℃温浴24h后经PR试剂分离结合和游离的标记抗原。用该法测定正常献血员150例和肝癌患者180例,并比较了肝癌患者介入治疗前后IGF-Ⅱ水平的变化。结果:该法测定范围为(0.1～24.3)ng/ml,最低检出值<0.1ng/ml,批内和批间误差<6.3%和10.0%。正常献血员血清IGF-Ⅱ水平(1.266±0.298)ng/ml明显低于肝癌各组患者(P<0.05);肝癌患者进行介入治疗3d后IGF-Ⅱ水平(4.263±1.411)ng/ml明显低于治疗前(4.916±1.24)ng/ml(P<0.05);介入治疗后3d与治疗后1个月(4.796±1.647)ng/ml水平相比无明显差异。结论:该法稳定、简便、特异、灵敏,适于检出人血清中的IGF-Ⅱ水平。
Objective To establish a highly specific and sensitive radioimmunoassay（RIA） for human insulin-like growth-factor- Ⅱ （IGF-Ⅱ）, and apply it to detect IGF-Ⅱ in human serum and culture medium. Methods Rabbits were immunized with recombined IGF-I1 connected with thyroglobulin（TG） for several times to obtain IGF-Ⅱantibody with high titer and specificity. Chlorozone-T method was used to prepare ,25 I-IGF-Ⅱ, and it was purified by Sephadex G-25. The reaction between antigen and antibody was procee- ded by one-step balancing method. They were incubated at 4℃ for 24h, and the binded or free labled antigen were separated by PR reagents. This assay was used to check the IGF- Ⅱ level in 150 samples from normal blood donors and 180 samples from patients with liver cancer, and compare change of IGF-Ⅱ level before and after interventional treatment in patients with liver cancer. Results The detective range of this assay was 0.1-24.3 ng/ml, the lowest detectable value was less than 0. I ng/ml, while the inter-/intra-assay deviation were less than 6.3% and 10.0% respectively. Serum IGF-Ⅱ of normal blood donors group （ 1. 266±0.298） ng/ml was signifi- candy lower than that in several liver cancer patients groups（P 〈0.05 ） ; while IGF-Ⅱ level of liver cancer patients without treatment （4.916±1.24）ng/ml was significantly higher than that in group treated with intervention 3 days （4. 263±1.411 ）ng/ml （P 〈 0.05 ）. However, no obviously disparity between group treated 3 days and group treated one month （4.796±1. 647 ng/ml）. Conclusion This IGF-Ⅱ radioimmunoassay is steady, convenient, specific and sensitive. Also, it can be used to detect IGF-Ⅱ level in human serum.
出处
《放射免疫学杂志》
CAS
2012年第3期241-242,共2页
Journal of Radioimmanology
关键词
人胰岛素样生长因子
放射免疫分析
肝癌
human insulin-like growth factor, radioimmunoassay（RIA）, liver cancer
