抗牛传染性鼻气管炎病毒囊膜gD糖蛋白的单链抗体在昆虫细胞中表达及活性检测

doi:10.16431/j.cnki.1671-7236.2017.08.036

Abstract

Abstract:

For preparation of bioactive single chain fragment variable (ScFv) which was targeted against envelope glycoprotein D (gD) of infectious bovine rhinotracheitis virus (IBRV),VH and VL genes were amplified from the cDNA of lymphocyte hybridoma, then VH and VL genes were integrated with a Linker by SOE-PCR,and the ScFv gene was cloned into the baculovirus vector pFB-LIC-Bse, which was transformed into Escherichia coli DH10Bac competent cells to construct a recombinant bacmid rBacmid-VH-VL. Finally,ScFv was expressed by transfected rBacmid-VH-VL into insect Sf9 cells, its molecular weight was about 30 ku. The result of Western blotting suggested that ScFv generated in Sf9 cells was specifically binds to His antibody,the protein also showed high affinity to gD of IBRV. The result of indirect immunofluorescence assay showed that ScFv could recognize IBRV which infected bovine kidney cells (MDBK). The study indicated that the recombinant ScFv was expressed in Sf9 cells, and could bind to gD of IBRV specifically. The result could provide the basis for the diagnosis and treatment of infection of IBRV.

Key words: infectious bovine rhinotracheitis virus (IBRV); glycoprotein D (gD); single chain fragment variable (ScFv) antibody; baculovirus expression system

CLC Number:

S852.65⁺9.1

WU Jing, XU Jian, HUANG Xiu-fen, SHEN Jun-jun, HE Hou-jun, LI Yong-qing. Expression and Biological Activity of Single Chain Antibody Against gD Glycoprotein of Infectious Bovine Rhinotracheitis Virus in Insect Cells[J]. , 2017, 44(8): 2475-2482.

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Expression and Biological Activity of Single Chain Antibody Against gD Glycoprotein of Infectious Bovine Rhinotracheitis Virus in Insect Cells

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