title: Mammalian cell display for rapid screening scFv antibody therapy.
abstract: Human antibodies are beginning to draw attention for use in immune gene therapy. The efficient generation of effective therapeutic monoclonal antibodies suitable for the treatment of cancers and infectious diseases would be enormously valuable. Antibody display methods are increasingly used to screen human monoclonal antibodies. Here we report the construction of a mammalian cell display method derived from a naive antibody repertoire, for which human single-chain variable fragments (scFv) have been transiently displayed on 293T cell surfaces based on a pDisplay vector. The sizes of the current pDisplay-scFv antibody repertoires have been estimated to be 0.74 × 10(7). An immunoblot assay confirmed the expression of the scFv antibody library. The subcellular distribution of ErbB3-scFv expression plasmid facilitated the display of ErbB3 scFv on the cell membrane surface and the efficiency of the display was evaluated by fluorescence-activated cell sorting. This method of mammalian cell display was verified by successfully screening ErbB3 scFv candidates. A published scFv control was used to confirm the feasibility of the ErbB3 scFv screening process. Three ErbB3 scFv candidates were produced and they were found to have affinity similar to the published scFv candidate. Thus, the present screening system provided an optimal alternative for rapid acquisition of a novel candidate scFv sequence to target genes with high affinity in vitro.
authors.affiliation: Beijing Institute of Radiation Medicine, Beijing 100850, China Anhui Medical University, Hefei 230032, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Zhang
authors.affiliation: Beijing Institute of Microbiology and Epidemiology, Beijing 100071, China.
authors.forename: Xiao'ai
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Beijing Institute of Radiation Medicine, Beijing 100850, China.
authors.forename: Qiang
authors.initials: Q
authors.lastname: Liu
authors.affiliation: Beijing Institute of Radiation Medicine, Beijing 100850, China.
authors.forename: Mengyi
authors.initials: M
authors.lastname: Li
authors.affiliation: Beijing Institute of Radiation Medicine, Beijing 100850, China.
authors.forename: Liucun
authors.initials: L
authors.lastname: Gao
authors.affiliation: Beijing Institute of Radiation Medicine, Beijing 100850, China.
authors.forename: Xin
authors.initials: X
authors.lastname: Gao
authors.affiliation: Beijing Institute of Radiation Medicine, Beijing 100850, China.
authors.forename: Shensi
authors.initials: S
authors.lastname: Xiang
authors.affiliation: Chinese PLA General Hospital, Tumor Center Laboratory, Beijing 100850, China.
authors.forename: Liangliang
authors.initials: L
authors.lastname: Wu
authors.affiliation: Beijing Institute of Radiation Medicine, Beijing 100850, China fujietiantian1@163.com bapklab@yahoo.com.
authors.forename: Jie
authors.initials: J
authors.lastname: Fu
authors.affiliation: Beijing Institute of Radiation Medicine, Beijing 100850, China fujietiantian1@163.com bapklab@yahoo.com.
authors.forename: Haifeng
authors.initials: H
authors.lastname: Song
mesh_terms: D001483:Base Sequence; D017931:DNA Primers; D005434:Flow Cytometry; D057809:HEK293 Cells; D006801:Humans; D007167:Immunotherapy; D057127:Single-Chain Antibodies / Q000276:immunology*
chemical_list: D017931:DNA Primers; D057127:Single-Chain Antibodies
keywords: FACS; erbb3; human embryonic kidney cell; human scfv antibody; mammalian cell display