丙型肝炎病毒抗原氨基酸序列多态性的质谱检测
Mass spectrometric detection of the amino acid sequence polymorphism of the hepatitis C virus antigen.
作者信息
Kaysheva A L, Ivanov Yu D, Frantsuzov P A, Krohin N V, Pavlova T I, Uchaikin V F, Konev V А, Kovalev O B, Ziborov V S, Archakov A I
机构信息
Institute of Biomedical Chemistry, Pogodinskaya St. 10, Moscow 119121 Russia.
Institute of Biomedical Chemistry, Pogodinskaya St. 10, Moscow 119121 Russia.
出版信息
J Virol Methods. 2016 Mar;229:86-90. doi: 10.1016/j.jviromet.2015.12.012. Epub 2016 Jan 7.
A method for detection and identification of the hepatitis C virus antigen (HCVcoreAg) in human serum with consideration for possible amino acid substitutions is proposed. The method is based on a combination of biospecific capturing and concentrating of the target protein on the surface of the chip for atomic force microscope (AFM chip) with subsequent protein identification by tandem mass spectrometric (MS/MS) analysis. Biospecific AFM-capturing of viral particles containing HCVcoreAg from serum samples was performed by use of AFM chips with monoclonal antibodies (anti-HCVcore) covalently immobilized on the surface. Biospecific complexes were registered and counted by AFM. Further MS/MS analysis allowed to reliably identify the HCVcoreAg in the complexes formed on the AFM chip surface. Analysis of MS/MS spectra, with the account taken of the possible polymorphisms in the amino acid sequence of the HCVcoreAg, enabled us to increase the number of identified peptides.
提出了一种考虑到可能的氨基酸替换来检测和鉴定人血清中丙型肝炎病毒抗原（HCVcoreAg）的方法。该方法基于在原子力显微镜芯片（AFM芯片）表面对目标蛋白进行生物特异性捕获和浓缩，并随后通过串联质谱（MS/MS）分析进行蛋白鉴定。通过使用表面共价固定有单克隆抗体（抗HCVcore）的AFM芯片，对血清样本中含有HCVcoreAg的病毒颗粒进行生物特异性AFM捕获。通过AFM对生物特异性复合物进行记录和计数。进一步的MS/MS分析能够可靠地鉴定在AFM芯片表面形成的复合物中的HCVcoreAg。考虑到HCVcoreAg氨基酸序列中可能的多态性，对MS/MS谱进行分析，使我们能够增加鉴定出的肽段数量。
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