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鲑鱼降钙素的灵敏时间分辨荧光免疫分析

Sensitive time-resolved fluoroimmunoassay of salmon calcitonin.

作者信息

Rong H, Tørring O, Sääf M, Sjöstedt U, Sjöberg H E, Bucht E

机构信息

Department of Molecular Medicine, Karolinska Hospital and Institute, Stockholm, Sweden.

出版信息

Clin Chem. 1994 Sep;40(9):1774-7.

PMID:8070090
Abstract

A two-site assay was developed by use of the "dissociation and enhancement lanthanide fluoroimmunoassay" (DELFIA) technique for determination of salmon calcitonin (SCT) in serum after administration to osteoporotic patients. Polyclonal antibodies were produced in rabbits immunized with SCT coupled to ovalbumin. After affinity purification, the antibodies were used both as immobilized capture antibodies and as Eu-chelate-labeled signal antibodies. A sensitive assay with a detection limit of 1.1 pmol/L was achieved, and no cross-reaction with human calcitonin was observed. The intra- and interassay CVs were < 12% (n = 10) and < 15% (n = 4), respectively. Analytical recovery of SCT added to serum was 91% +/- 3% (mean +/- SD, n = 4). SCT was measurable in all the samples from eight osteoporotic patients after subcutaneous SCT administration. We conclude that this new sensitive and specific two-site DELFIA can reliably measure SCT in serum.

摘要

采用“解离增强镧系元素荧光免疫分析”(DELFIA)技术开发了一种双位点分析方法,用于测定骨质疏松症患者给药后血清中的鲑鱼降钙素(SCT)。用与卵清蛋白偶联的SCT免疫家兔制备多克隆抗体。经亲和纯化后,这些抗体既用作固定化捕获抗体,也用作铕螯合物标记的信号抗体。该分析方法灵敏度高,检测限为1.1 pmol/L,且未观察到与人降钙素的交叉反应。批内和批间变异系数分别<12%(n = 10)和<15%(n = 4)。添加到血清中的SCT的分析回收率为91%±3%(平均值±标准差,n = 4)。皮下注射SCT后,八名骨质疏松症患者的所有样本中均可检测到SCT。我们得出结论,这种新的灵敏且特异的双位点DELFIA能够可靠地测定血清中的SCT。

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