Abstract: Objective To construct a personalized full-length fully human antibody mammalian display library for children with
systemic lupus erythematosus (SLE). Methods The total RNA was isolated from the PBMCs of SLE children. The heavy chain
variable region and kappa light chain (VH and LCκ) of the antibody genes were amplified by RT-PCR and inserted into the
pDGB-HC-TM vector separately to construct the heavy chain and light chain libraries. The library DNAs were transfected into
293T cells and the expression of full-length fully human antibody on the surface of 293T cells was analyzed by flow cytometry.
Results Using 0.8 μg total RNA as the template, the VH and LCκ were amplified and the full-length fully human antibody
mammalian display library was constructed. The VH and LCκ gene libraries had a size of 9.4×104 and 8.4×104, respectively.
Sequence analysis of 10 clones randomly selected from the VH and LCκ gene libraries each showed that 8 heavy chain clones
and 7 light chain clones contained correct open reading frames, and flow cytometry demonstrated that all the 15 clones express
full-length antibodies on 293T cell surfaces. 293T cells co-transfected with the VH and LCκ gene libraries expressed the
full-length antibodies on the cell surface. Conclusion The personalized full-length fully human antibody library for SLE
children constructed allows display of the full-length antibodies on mammalian cell surfaces, thus providing a valuable
platform for analyzing the autoantibodies, their etiological role, and their clinical implications in SLE.