Abstract: Objective To construct full-length human bladder cancer-specific antibody libraries for efficient display of
full-length antibodies on the surface of mammalian cells. Methods The total RNA was isolated from peripheral blood
mononuclear cells from patients with bladder cancer. The repertoires of IgG1 heavy chain variable region (VH) and Kappa
light chain were amplified by RT-PCR using specific primers. The antibody genes were inserted into the vector pDGB-HC-TM
to construct the bladder-cancer-specific antibody libraries of heavy chains and light chains. Ten clones from each library were
randomly picked for gene sequencing and transient transfection into FCHO cells to analyze antibody display on mammalian
cell surface by flow cytometry after staining with corresponding fluorescent labeled antibodies. Results The libraries of
bladder-cancer-specific antibody heavy chain (IgG1) and light chain (LCκ) were successfully constructed. Seven out of the 10
clones randomly selected from the heavy chain library and 9 out of the 10 clones from the light chain library showed correct
open reading frame, coding for 7 unique VH and 9 unique LCk. The combinatory library size reached 3.32×1011 [(1.7×106×70%) ×
(3.1 × 105 × 90%)]. Conclusion We have successfully constructed a full-length human bladder-cancer-specific antibody library
with a combinatory diversity of 3.32 × 1011 based on mammalian display technology, which can be used for screening
monoclonal antibodies against bladder-cancer-associated antigens.