Abstract: Objective To construct dengue virus-specific full-length fully human antibody libraries using mammalian cell
surface display technique. Methods Total RNA was extracted from peripheral blood mononuclear cells (PBMCs) from
convalescent patients with dengue fever. The reservoirs of the light chain and heavy chain variable regions (LCκ and VH) of
the antibody genes were amplified by RT-PCR and inserted into the vector pDGB-HC-TM separately to construct the light
chain and heavy chain libraries. The library DNAs were transfected into CHO cells and the expression of full-length fully
human antibodies on the surface of CHO cells was analyzed by flow cytometry. Resluts Using 1.2 μg of the total RNA isolated
from the PBMCs as the template, the LCκ and VH were amplified and the full-length fully human antibody mammalian
display libraries were constructed. The kappa light chain gene library had a size of 1.45×104 and the heavy chain gene library
had a size of 1.8×105. Sequence analysis showed that 8 out of the 10 light chain clones and 7 out of the 10 heavy chain clones
randomly picked up from the constructed libraries contained correct open reading frames. FACS analysis demonstrated that
all the 15 clones with correct open reading frames expressed full-length antibodies, which could be detected on CHO cell
surfaces. After co-transfection of the heavy chain and light chain gene libraries into CHO cells, the expression of full-length
antibodies on CHO cell surfaces could be detected by FACS analysis with an expressible diversity of the antibody library
reaching 1.46 × 109 [(1.45 × 104 × 80%) × (1.8 × 105 × 70%)]. Conclusion Using 1.2 μg of total RNA as template, the LCκ and VH
full-length fully human antibody libraries against dengue virus have been successfully constructed with an expressible
diversity of 109.