葡萄藤病毒B重组抗体（单链抗体片段）的分离

Isolation of recombinant antibodies (scFvs) to grapevine virus B.

作者信息

Saldarelli P, Keller H, Dell'Orco M, Schots A, Elicio V, Minafra A

机构信息

Istituto di Virologia Vegetale del CNR, Sezione di Bari, Italy.

出版信息

J Virol Methods. 2005 Mar;124(1-2):191-5. doi: 10.1016/j.jviromet.2004.11.021. Epub 2004 Dec 28.

DOI:10.1016/j.jviromet.2004.11.021

PMID:15664068

Abstract

A panel of 15 recombinant single chain antibodies (scFv) specific to grapevine virus B (GVB) were recovered from a human combinatorial scFv antibody library using the phage display technique against purified virus particles. Two selected scFv-encoding genes were expressed in recombinant Escherichia coli cells as dimeric antibodies. Successful detection of GVB in tissues of herbaceous hosts and grapevine was obtained in a direct binding assay using dimeric scFvs. This reagent was also shown to substitute efficiently for a GVB polyclonal serum in standard DAS-ELISA test used routinely for diagnosis.

摘要

利用噬菌体展示技术，针对纯化的病毒颗粒，从人源组合单链抗体（scFv）文库中筛选出15种针对葡萄扇叶病毒（GVB）的重组单链抗体。两个选定的编码scFv的基因在重组大肠杆菌细胞中表达为二聚体抗体。在使用二聚体scFv的直接结合试验中，成功检测到草本寄主和葡萄组织中的GVB。在常规用于诊断的标准双抗夹心酶联免疫吸附测定（DAS-ELISA）试验中，该试剂也被证明能有效替代GVB多克隆血清。