Sensing HIV related protein using epitope imprinted hydrophilic polymer coated quartz crystal microbalance
Sensing HIV related protein using epitope imprinted hydrophilic polymer coated quartz crystal microbalance
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使用表位印迹亲水性聚合物涂层石英晶体微天平感测 HIV 相关蛋白
DOI：
10.1016/j.bios.2011.11.008
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发表时间：
2012-01-15
影响因子：
12.6
通讯作者：
Chen, Guo-Nan
中科院分区：
文献类型：
--
作者：
Lu, Chun-Hua;Zhang, Yan;Chen, Guo-Nan
我们开发了一种基于表位印迹技术的仿生传感器，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）。 gp41是HIV-1的跨膜蛋白，在病毒与感染细胞之间的膜融合中发挥重要作用。它是判断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，并在模板存在下在石英晶体微天平（QCM）芯片的表面上聚合，模板是一种具有35个氨基酸残基的合成肽，类似于gp41的残基579-613。该过程导致在 QCM 芯片上接枝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，而且能够特异性结合相应的gp41蛋白。通过Scatchard分析计算出模板肽的MIP解离常数(K-d)为3.17nM，与单克隆抗体的解离常数相似。使用由此产生的基于 MIP 的仿生传感器定量地实现了 gp41 的直接检测。 gp41的检测限为2 ng/mL，与报道的ELISA方法相当。此外，通过评估人尿液样本中gp41的检测来检验传感器的实际分析性能，结果令人满意。 (C) 2011 Elsevier B.V. 保留所有权利。
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. (C) 2011 Elsevier B.V. All rights reserved.