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利用表位印迹亲水聚合物涂层石英晶体微天平检测 HIV 相关蛋白。

Sensing HIV related protein using epitope imprinted hydrophilic polymer coated quartz crystal microbalance.

机构信息

The Key Lab of Analysis and Detection Technology for Food Safety of the MOE, Fujian Provincial Key Laboratory of Analysis and Detection Technology for Food Safety, College of Chemistry and Chemical Engineering, Fuzhou University, Fuzhou 350002, PR China.

出版信息

Biosens Bioelectron. 2012 Jan 15;31(1):439-44. doi: 10.1016/j.bios.2011.11.008. Epub 2011 Nov 15.

Abstract

We have developed a biomimetic sensor for the detection of human immunodeficiency virus type 1 (HIV-1) related protein (glycoprotein 41, gp41) based on epitope imprinting technique. gp41 is the transmembrane protein of HIV-1 and plays an important role in membrane fusion between viruses and infected cells. It is an important index for determining the extent of HIV-1 disease progression and the efficacy of therapeutic intervention. In this work, dopamine was used as the functional monomer and polymerized on the surface of quartz crystal microbalance (QCM) chip in the presence of template, a synthetic peptide with 35 amino acid residues, analogous to residues 579-613 of the gp41. This process resulted in grafting a hydrophilic molecularly imprinted polymer (MIP) film on the QCM chip. QCM measurement showed that the resulting MIP film not only had a great affinity towards the template peptide, but also could bind the corresponding gp41 protein specifically. The dissociation constant (K(d)) of MIP for the template peptide was calculated to be 3.17 nM through Scatchard analysis, which was similar to those of monoclonal antibodies. Direct detection of the gp41 was achieved quantitatively using the resulting MIP-based biomimetic sensor. The detection limit of gp41 was 2 ng/mL, which was comparable to the reported ELISA method. In addition, the practical analytical performance of the sensor was examined by evaluating the detection of gp41 in human urine samples with satisfactory results.

摘要

我们基于表位印迹技术开发了一种用于检测人类免疫缺陷病毒 1 型(HIV-1)相关蛋白(糖蛋白 41,gp41)的仿生传感器。gp41 是 HIV-1 的跨膜蛋白,在病毒与感染细胞之间的膜融合中起着重要作用。它是确定 HIV-1 疾病进展程度和治疗干预效果的重要指标。在这项工作中,多巴胺被用作功能单体,并在模板(与 gp41 的残基 579-613 类似的 35 个氨基酸残基的合成肽)存在的情况下聚合在石英晶体微天平(QCM)芯片的表面上。这导致在 QCM 芯片上接枝了一层亲水性分子印迹聚合物(MIP)薄膜。QCM 测量表明,所得的 MIP 薄膜不仅对模板肽具有很大的亲和力,而且还可以特异性地结合相应的 gp41 蛋白。通过 Scatchard 分析计算出 MIP 对模板肽的解离常数(Kd)为 3.17 nM,与单克隆抗体相当。通过使用所得的基于 MIP 的仿生传感器可以定量地直接检测 gp41。gp41 的检测限为 2 ng/mL,与报道的 ELISA 方法相当。此外,通过评估人尿液样品中 gp41 的检测,检验了传感器的实际分析性能,结果令人满意。

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