摘要
用夹心法建立铁蛋白(FER)时间分辨荧光免疫分析法(TRFIA),测定血清FER的含量。以FER单克隆抗体(McAb)806#包被板,双功能螯合剂异氰酸苄基二乙烯三胺四乙酸络合Eu3+及标记FERMcAb803#,发光增强系统为以β二酮体为主的增强液。采用双对数函数数据处理程序,方法的批内和批间CV分别为2.2%和3.7%,平均回收率为118%,灵敏度为0.5μg/L,可测范围为0.51000μg/L。ED20、ED50和ED80分别为27.8μg/L、88.5μg/L和247μg/L。Eu3+FERMcAb于-30℃保存至少4个月,免疫反应性基本无损失,同批试剂连续分析4个月结果稳定。本法与PE公司的FERTRFIA试剂盒所得样品的测定值的相关系数为0.98。本文建立的FERTRFIA法,是一种适用于人群贫血普查和各种肝脏疾病及肿瘤的联合诊断的良好方法,分析范围、灵敏度和稳定性均较理想,值得推广应用。
A two-site time-resolved fluoroimmunoassay (TRFIA) for ferritin(FER) based on the direct sandwich technique was developed. The McAb(806#) was directed against a specific antigenic site on the FER. The europium-labeled McAb(803#) which directed against a diffe- rent antigenic site on the FER was prepared by using europium-chelated N_2-(p-isothiocyanatobenzyl)-diethylenetriamine-N,N,N-tetraacetic acid. The luminescent enhancement system was an enhancement solution containing mainly 2-naphthoyltrifluoroacetone. The intra-and inter-assay CV of the TRFIA were 2.2% and 3.7%, respectively, the recovery rate was 118%, and the sensitivity of FER was 0.5 μg/L. The FER-TRFIA provided a linear response from 0.5 to 1000μg/L FER with ED_ 80 of 27.8μg/L, ED_ 50 of 88.5μg/L and ED_ 20 of 247μg/L. The europium-labeled McAb(803#) was stable at least for four months at -30℃. The results of the TRFIA with same reagents were stable for four months. Therefore, the FER-TRFIA established here is a suitable method for detecting the serum levels of FER for diagnosis of anemia, hepatic disease and tomour.
出处
《标记免疫分析与临床》
CAS
2005年第2期104-106,113,共4页
Labeled Immunoassays and Clinical Medicine
