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基于介孔硅和胶体金纳米粒子的新型敏感免疫层析检测方法的建立。

Development of a new sensitive immunostrip assay based on mesoporous silica and colloidal Au nanoparticles.

机构信息

Endocrinology and Metabolism Research Center, Tehran University of Medical Sciences, P.O. Box 14395/1179 Tehran, Islamic Republic of Iran.

出版信息

Mol Biol Rep. 2012 Feb;39(2):1253-9. doi: 10.1007/s11033-011-0856-5. Epub 2011 May 21.

Abstract

A new competitive immunostrip assay was developed to detect human serum albumin (HSA) in urine sample with use of conjugated monoclonal antibody gold nanoparticles (mAb-AuNPs) and mobile crystalline material (MCM)-41-HSA bioconjugate. To prepare the immunostrip, the colloidal AuNPs with an average particle diameter of 20 nm, was synthesized, labeled with antibody and applied on the conjugate pad as the detection reagent. Then, HSA was attached to the MCM-41 mesoporous nanoparticles and immobilized to a nitrocellulose membrane as the test line. In the optimized investigational conditions, the immunostrip could detect HSA in a high linear range (from 1 to 200 μg/ml) and low detection limit (ng/ml). The reliability of the testing procedure was examined by performing the immunostrip test with 30 urine samples and comparing the results with those obtained via immunoturbidimetry. The immunostrip was adequately sensitive and accurate for a rapid screening of HSA in the urine. This new strategy for competitive immunostrip design can be used and developed for other antigen based immunostrip assay.

摘要

一种新的竞争免疫层析检测法被开发出来,用于检测尿液样本中的人血清白蛋白(HSA),该方法使用了结合的单克隆抗体金纳米粒子(mAb-AuNPs)和可移动结晶材料(MCM)-41-HSA 生物缀合物。为了制备免疫层析检测条,合成了平均粒径为 20nm 的胶体金纳米粒子,并用抗体进行标记,并将其作为检测试剂应用于结合垫上。然后,将 HSA 附着到 MCM-41 介孔纳米粒子上,并将其固定在硝酸纤维素膜上作为测试线。在优化的研究条件下,免疫层析检测条可以在高线性范围内(从 1 到 200μg/ml)和低检测限(ng/ml)下检测 HSA。通过对 30 个尿液样本进行免疫层析检测,并将结果与免疫比浊法的结果进行比较,检验了检测程序的可靠性。该免疫层析检测条对尿液中 HSA 的快速筛选具有足够的灵敏度和准确性。这种竞争免疫层析设计的新策略可用于和开发其他基于抗原的免疫层析检测。

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