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目的 用葡萄球菌蛋白A(StaphylococcalProteinA ,SPA)制备固相抗体并应用于放射免疫分析。方法 预先将SPA用物理吸附法包被到聚苯乙烯塑料试管上 ,利用SPA 抗体 放射性标记抗原结合模式 ,通过测定管壁所结合放射性大小来研究SPA抗体固化技术。制备固相甲状腺素抗体 (T4 Ab)用于T4放射免疫分析并与对照试剂盒对照。结果 SPA包被 :1μg /ml于pH9 6碳酸盐缓冲液 ;T4 Ab包被 :1∶30 0 0 0于pH8 4磷酸盐缓冲液包被 ,最佳包被管结合率 45 6 % ;批内变异 3 5 % ,批间变异 6 6 % ;连续洗涤 8次 ,平均每次洗涤后结合率下降 <0 5 % ;固相抗体管 4℃保存 2个月 ,最大结合率不变 ;与对照试剂盒比较测定 43份样品所测值差异无显著性 (t=- 0 717,P =0 494)。结论 SPA固化抗体简单、均一、稳定、重复性好 ,可以用于放射免疫分析。
Objective To prepare solid-phase antibody with Staphylococcal Protein A (SPA) and apply it to radioimmunoassay. Methods The SPA was pretreated by physical adsorption on a polystyrene plastic tube and the SPA antibody immobilization technique was studied by measuring the radioactivity bound to the tube wall by using SPA antibody radiolabeled antigen binding mode. Solid phase thyroxine antibodies (T4 Ab) were prepared for T4 radioimmunoassay and control against the control kit. Results The results of the SPA coating were: 1μg / ml at pH9 6 carbonate buffer; T4 Ab coating: 1:30 0 0 0 coated with phosphate buffer pH8 4, the best package tube binding rate 45 6%; batch The intra-assay variation was 35%, and the inter-assay variation was 6 6%. The serial washes were performed 8 times, with an average of <0 5% reduction in the binding rate after each wash. The solid phase antibody tubes were kept at 4 ° C for 2 months, There was no significant difference between the test samples and the 43 samples (t = - 0 717, P = 0 494). Conclusion SPA curing antibody is simple, uniform, stable, reproducible, and can be used for radioimmunoassay.