猪伪狂犬病病毒gE蛋白单链抗体的制备及鉴定

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中图分类号：S852.659.1 文献标志码：A 文章编号：0366-6964（2026）01-0317-10

Abstract：Since 2Ol1，the new outbreak of pseudorabies virus（PRV） variant strain in China has greatly increased the detection workload of the virus. At present，the widely use of blocking ELISA kits has raised the greater demand for monclonal antibodies.However，hybridoma cells often lose antibody secretion capacity during passages. This study aimed to prepare single-chain fragment variable with good binding activity on the basis of monoclonal antibody and achieve large-scale production via prokaryotic expression system，providing a foundation for novel detection methods. The prokaryotically expressed PRV gE protein was used to immunize mice. Positive hybridoma cells stably secreting anti-PRV gE mabs were obtained through cell fusion and screening. The cDNA of the positive hybridoma cells was used as a template to amplify the variable region sequence of antibody，and the scFv gene was constructed by linking the variable regions of heavy and light chain with a Linker peptide. The scFv gene was cloned into pET-28a plasmid for prokaryotic expression to produce the recombinant antibody. A monoclonal antibody 7B7 specific to the gE protein of PRV was successfully screened. Using the cDNA of 7B7 hybridoma cels as a template，a single chain fragment variable capable of specifically reacting with PRV gE was successfully prepared which can be expressed by prokaryotic expression system，providing a basis for the development of new detection methods for PRV.

Keywords：pseudorabies virus；gE protein；single chain fragment variable Corresponding author： HE Qigai，E-mail： he628@mail.hzau.edu. cn

猪伪狂犬病（pseudorabies，PR）是由猪伪狂犬病病毒（pseudorabiesvirus，PRV）引起的多种家畜及野生动物感染的急性、热性传染病[1]。（剩余15003字）