抗辛诺柏汉坦病毒核衣壳蛋白单链抗体片段的筛选与鉴定

Selection and characterization of scFv antibodies against the Sin Nombre hantavirus nucleocapsid protein.

作者信息

Velappan Nileena, Martinez Jennifer S, Valero Rosa, Chasteen Leslie, Ponce Liana, Bondu-Hawkins Virginie, Kelly Craig, Pavlik Peter, Hjelle Brian, Bradbury Andrew R M

机构信息

Los Alamos National Laboratory, TA-43, HRL-1, MS M888, Los Alamos NM 87545, USA.

出版信息

J Immunol Methods. 2007 Apr 10;321(1-2):60-9. doi: 10.1016/j.jim.2007.01.011. Epub 2007 Feb 8.

DOI:10.1016/j.jim.2007.01.011

PMID:17336997

Abstract

Rodent-borne hantaviruses cause hemorrhagic fever with renal syndrome (HFRS) in the old world and hantavirus cardio-pulmonary syndrome (HCPS) in the new. Most cases of HCPS in North America are caused by Sin Nombre Virus (SNV). Current viral detection technologies depend upon the identification of anti-viral antibodies in patient serum. Detection of viral antigen may facilitate earlier detection of the pathogen. We describe here the characterization of two single-chain Fv antibodies (scFvs), selected from a large naïve phage antibody library, which are capable of identifying the Sin Nombre Virus nucleocapsid protein (SNV-N), with no cross reactivity with the nucleocapsid protein from other hantaviruses. The utility of such selected scFvs was increased by the creation of an scFv-alkaline phosphatase fusion protein which was able to directly detect virally produced material without the need for additional reagents.

摘要

啮齿动物传播的汉坦病毒在旧世界引起肾综合征出血热（HFRS），在新世界引起汉坦病毒心肺综合征（HCPS）。北美大多数HCPS病例由辛诺柏病毒（SNV）引起。目前的病毒检测技术依赖于在患者血清中鉴定抗病毒抗体。病毒抗原的检测可能有助于更早地检测病原体。我们在此描述了从一个大型天然噬菌体抗体库中筛选出的两种单链Fv抗体（scFvs）的特性，它们能够识别辛诺柏病毒核衣壳蛋白（SNV-N），且与其他汉坦病毒的核衣壳蛋白无交叉反应。通过创建scFv-碱性磷酸酶融合蛋白提高了此类筛选出的scFvs的实用性，该融合蛋白能够直接检测病毒产生的物质，无需额外试剂。