Preparation and Epitope Analysis of Monoclonal Antibody against African Swine Fever Virus D205R Protein
LIN Zhizhao
ZHAO Yanyan
REN Haojie
SHI Saiyan
HAN Shichong
HE Wenrui
WAN Bo
ZHANG Yuhang
ZHANG Gaiping
Abstract：African swine fever(ASF)is a lethal infectious disease,and few vaccines or drugs were available to prevent or control ASF to date.In order to provide important materials for the diagnosis of African swine fever virus(ASFV),the ASFV recombinant D205R protein was expressed by E.coli system,a monoclonal antibody(mAb)against D205R protein was prepared,and antigenic epitope of mAb was identified.The results showed that the pET32a-D205R expression plasmid was successfully constructed,and the recombinant D205R protein with a size of about 44 ku was purified.Western blot assay showed that the recombinant D205R protein reacted specifically with ASFV-positive serum and had good immunogenicity.By hybridoma cell fusion and screening,mAb 19A5 was obtained.Western blot and indirect immunofluorescence assays showed that mAb 19A5 specifically recognized eukaryotically expressed recombinant D205R protein,and detected wild-type D205R protein.Alanine scanning identified 167-SDPPVVWLGGRPGD-180 as the antigenic epitope of mAb 19A5,and S167,W173,L174,G175,P178,and D180 were the key amino acids that binded to mAb 19A5.Homologous and structural analysis revealed that the antigenic epitope was highly conserved and located on the surface of the protein,which was a linear epitope.In conclusion,mAb 19A5 was successfully prepared and the antigenic epitope recognized by mAb 19A5 was identified.
Keywords：African swine fever virusD205R proteinMonoclonal antibodyAntigenic epitope
Publication Date：2025-02-15
Online Publishing Date：2026-09-12(First online date of this platform, not the publication date of the document)
Pages：7( 124-130 )