共价结合和物理吸附制备固相抗体在RIA技术应用中的对比分析

Study on the Technical Parameters of Two Different Systems of RIA Performed with Solid-Phase Antibody Test Tubes Prepared with Magnetic Microparticle Covalence Conjagation or Conventional Physical Absorption

目的:探讨异硫氰酸荧光素(FITC)-抗FITC磁性微粒子系统与试管固相包被系统制备固相抗体在RIA应用中的方法学技术参数。方法:以FITC标记抗体,抗FITC抗体共价结合磁性微粒子为分离剂,与试管固相包被法,用相同的校准品、125I标记物通过检测FT3、sTSH进行技术研究及样本对比分析。结果:FT3两种方法的灵敏度和精密度分别为:0.18pmol/L、0.43pmol/L,批内变异(n=10):8.96%、16.26%,批间变异(n=10):15.26%、18.83%。sTSH两种方法的灵敏度和精密度分别为:0.06μIU/ml、0.04μIU/ml,批内变异(n=10):7.6%、6.92%,批间变异(n=10):8.55%、14.23%。FT3磁性微粒子法测值与PR法测值r=0.9825,FT3试管固相包被法测值与PR法测值r=0.9102;sTSH磁性微粒子法测值与试管固相包被法测值r=0.9987。结论:FITC磁性微粒子系统应用于FT3、sTSH检测,降低反应时间,共价结合提高均一性。

Objective To investigate a new method of preparation of solid - phase antibody with fluroreseein isothioeynate （FITC） -anti FITC magnetic nanopartieles system （for FT3 and TSH）. Methods FT3 and TSH monoelonal antibody IgG was labelled with FITC. Anti - FITC magnetic mieroparfieles was prepared and conjugated with the FITC labelled antibody to form the solid - phase coated test tube for RIA. Solid - phase test tube prepared with the conventional physical absorption method was also used for RIA and the technical parameters of the two systems were compared. Results For FT3 , the sensitivity was 0. 18pmol/L with the new method and 0.43pmol/L with the conventional method. Other parameters were ; intra - CV, 8.96% vs 16.26% ; inter - CV, 15.25% va 18.83%, correlation with PR method r = 0.9825 vs r = 0. 9102. For TSH, sensitivity was 0.06μU/ml vs 0.041μIU/ml, intra - CV, 7.6% vs 6.92%, inter- CV, 8.55% vs 14.23%, correlation between the new and conventional method, r = 0. 9987. TSH RIA was especially rapid with the new technic： 79min vs 190min. Conclusion For FT3 and TSH R/A, the new method takes much less time with increased homogeniety.