Cell display library for gene cloning of variable regions of human antibodies to hepatitis B surface antigen

Development of a novel mammalian cell surface antibody display platform

“…19 Ho et al have described a system to display single chain anti-CD22 Fv fragments on 293T cells. 21 Both groups have successfully screened for clones expressing antibodies with antigen specificity 19 or higher affinity. 21 Standard transfection or electroporation methods, however, will typically introduce multiple copies of vectors into each cell.…”

“…It also becomes extremely difficult to compare precisely the affinity and expression on the mammalian cell surface and isolate the desired cell clones efficiently. It has been realized that, in both systems mentioned, 19,21 the calculated diversity required for selection of clones with better affinity or antigen specificity is about one thousand or less. This relative abundance of the desired populations in a library facilitates the screening in randomly integrated or transiently transfected systems.…”

“…Researchers have applied multiple strategies in the effort to develop mammalian display technology, including using different mammalian cell types, e.g., COS cells, lymphoma-derived B cells, 293T cells, BHK cells and different delivery vehicles, e.g., plasmids, retroviral vectors, vaccinia virus and sindbis virus, to display peptides, antibody fragments or full-length antibodies. [19][20][21][22][23][24][25][26] Most of these systems, however, display multiple copies of antibodies with different specificities on a single cell surface, making it difficult to identify and isolate antibodies with a desired property. These limitations significantly hamper the application of mammalian display technology in the development of therapeutic antibodies.…”

A novel strategy for rapid construction of libraries of full-length antibodies highly expressed on mammalian cell surfaces

“…During the past 10 years, many scientists have exercised considerable effort in the attempt to develop mammalian cell surface display technology. It is therefore desirable to develop technology that can display full-length antibodies, especially human antibodies, on mammalian cell surfaces [11,12]. Some techniques have fused a TM domain at the 3 0 -end of the antibody heavy chain (HC) to form the HC-TM.…”

“…Some techniques have fused a TM domain at the 3 0 -end of the antibody heavy chain (HC) to form the HC-TM. Co-expressing this HC-TM with a light chain (LC), the full-length antibody could be displayed on the cell surface [12]. Coupled with fluorescence activated cell sorting (FACS), some antigen-specific antibodies were successfully screened and identified from mammalian antibody libraries displayed on the cell surfaces [11][12][13].…”