【摘要】 目的制备地高辛免疫试剂盒所需的特异抗体和酶标抗原。方法化学合成地高辛(Dig)、牛血清白蛋白(BSA)结合物(Dig-BSA),形成完全抗原;依据动物体的免疫反应规律安排免疫日程,制备高质量的抗地高辛特异性抗体(Dig-Ab)。化学合成酶标记抗原,将地高辛抗体和酶标抗原应用于地高辛血浓度监测的免疫脂质体溶破法(LILA)、酶联免疫吸附试验(ELISA)和化学发光酶免疫法(CLEIA)测定。结果抗体效价达1:1.8×10~6放射免疫法(RIA),亲合系数为2.67×10~9,与人血清内源物质无交叉反应。三种免疫方法的灵敏度、线性、回收率、精密度分别为0.2～0.06μg/L、10μg/L、89.3%～110.5%、2.4%～8.8%(批内)、6.0%～18.8%(批间),与荧光偏振法和 RIA 法有良好相关。结论制备的抗体和酶标抗原可用于地高辛的免疫试剂盒。更多还原

【Abstract】 Objective To prepare the specific antibody against digoxin(anti-Dig)and enzyme la- beled digoxin for digoxin immunoassay kits.Methods Specific antibodies against digoxin(anti-Dig) were obtained by immunizing rabbits with chemically conjugated Dig-BSA.The anti-DIG and enzyme labeled digoxin were applied to three different kinds of immunoassay for determining serum digoxin lev- els.Results The anti-Dig antibodies with high titer(as high as 1:1.8×10~6 by RIA),high affinity (as high as 2.67×10~9)and high specificity(no cross-reactions with endogenous substances)were ob- tained.In the three kinds of immunoassay(ELISA,LILA and CLEIA)the detecting limit was 0.2～ 0.06μg/L,within run and run-to-run CVs was 2.4%～8.8% and 6.0%～18.8% respectively,and the recovery rate was 89.3%～110.5%.The correlation with RIA and FPIA is excellent,r= 0.9200～0.9400.Conclusions The anti-DIG and the enzyme labeled digoxin can be used was digox- in immunoassay kits.更多还原