Screening and identification of human antibodies against dengue virus (DENV) envelope protein
GAO Bo1, 2, ZHENG Hui2, LI Shu2, XIE Lian-hui3
1.School of Life Science, Fujian Agriculture and Forestry University,Fuzhou 350001 China; 2.Fujian International Travel Health Center, Fuzhou 350001 China; 3.Institute of Plant Virology,Fujian Agriculture and Forestry University, Fuzhou 350001,China
Abstract：We amied to establish a method for rapid screening of human antibodies against dengue virus(DENV) capsids from peripheral blood, and to screen specific antibodies from peripheral blood memory B cells of virus-infected patients for qualitative analysis. By flow cytometry, DENV envelope protein-specific memory B cells in peripheral blood were single-cell sorted. The gene sequence of antibody was amplified using efficient single-cell PCR. We determined the binding activity and neutralization potency of recombinant full-length antibodies. Six human monoclonal antibodies recognizing DENV-1 envelope protein were successfully screened, and four of them had strong binding activity and neutralizing activity. DENV-specific memory B cells were successfully sorted by flow cytometry, and DENV-specific human monoclonal antibodies were efficiently obtained using single-cell PCR. The results revealed that it is hopeful to obtain DENV-specific antibody with broadly neutralizing activity for the prevention and treatment of dengue fever.
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