大鼠淋巴细胞培养上清液总IgG含量ELISA检测方法的建立  

Establishment of ELISA for Total IgG Content of Rat Lymphocyte Culture Supernatant

作　　者：单纯筱[1] 王宇[1] 尹磊淼[1] 徐玉东[1] 魏颖[1] 冉君[1] 刘晓燕[1] 刘奇[1] 杨永清[1] 

机构地区：[1]上海中医药大学上海市针灸经络研究所,上海200030

出　　处：《现代生物医学进展》2012年第26期5015-5019,共5页Progress in Modern Biomedicine

基　　金：国家自然科学基金(81001548;81173341;81173332;30873286);上海市教委和上海市教育发展基金会"晨光计划"资助项目(10CG45);上海市卫生局青年基金(2009Y096);国家中医药管理局;上海市重点学科建设项目(S30304)

摘　　要：目的:建立一种定量测定大鼠淋巴细胞培养上清液中总IgG(免疫球蛋白G)含量的双抗体夹心ELISA(酶联免疫吸附试验)检测方法。方法:用方阵实验确定包被抗体、检测抗体的最佳工作浓度;绘制标准曲线,确定线性范围;评价标准曲线的可重复性、精密度和可应用性。结果:包被抗体和检测抗体的最佳效价分别为2μg/ml和1:4000稀释;检测的线性范围为0.25-16ng/ml。经方法学评价,可重复性和精密度较高,应用性较强。结论:该方法灵敏度高,重复性好,可作为科研过程中检测大鼠淋巴细胞培养上清液总IgG含量的一种精确、方便、可靠的方法。Objective： To establish a quantitative double-antibody sandwich ELISA detecting method for total IgG content of cultured rat-lymphocyte supematant. Methods： Detect the optimal concentrations of coated antibody and enzyme-antibody with a matrix experiment. Construct a calibration curve and determine the linear rang. Evaluation repeatability, precision and applicability of the calibration curve. Results： The optimal concentration of coated antibody is 2 ug/ml and the optimal dilution ratio of enzyme-antibody is 1：4000. The linear rang of calibration curve is 0.25-16 ng/mL. The repeatability, precision and applicability are excellent. Conclusion： This ELISA detecting method is convenient and reliable to detect total IgG content of cultured rat-lymphocyte supernatant.

关 键 词：大鼠淋巴细胞培养上清液 IGG 双抗体夹心ELISA 

分 类 号：Q95-33[生物学—动物学] R446[医药卫生—诊断学]