title: Development of a novel mammalian cell surface antibody display platform.
abstract: Antibody display systems have been successfully applied to screen, select and characterize antibody fragments. These systems typically use prokaryotic organisms such as phage and bacteria or lower eukaryotic organisms, such as yeast. These organisms possess either no or different post-translational modification functions from mammalian cells and prefer to display small antibody fragments instead of full-length IgGs. We report here a novel mammalian cell-based antibody display platform that displays full-length functional antibodies on the surface of mammalian cells. Through recombinase-mediated DNA integration, each host cell contains one copy of the gene of interest in the genome. Utilizing a hot-spot integration site, the expression levels of the gene of interest are high and comparable between clones, ensuring a high signal to noise ratio. Coupled with fluorescence-activated cell sorting (FACS) technology, our platform is high throughput and can distinguish antibodies with very high antigen binding affinities directly on the cell surface. Single-round FACS can enrich high affinity antibodies by more than 500 fold. Antibodies with significantly improved neutralizing activity have been identified from a randomly mutagenized library, demonstrating the power of this platform in screening and selecting antibody therapeutics.
authors: Protein Science, Amgen Inc., Thousand Oaks, CA, USA.
authors: Chen
authors: C
authors: Zhou
authors: Frederick W
authors: FW
authors: Jacobsen
authors: Ling
authors: L
authors: Cai
authors: Qing
authors: Q
authors: Chen
authors: Weyen David
authors: WD
authors: Shen
mesh_terms: D000818:Animals; D000906:Antibodies / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D000915:Antibody Affinity / Q000276:immunology; D015153:Blotting, Western; D016466:CHO Cells; D002462:Cell Membrane / Q000378:metabolism*; D006224:Cricetinae; D003412:Cricetulus; D005434:Flow Cytometry; D057809:HEK293 Cells; D006801:Humans; D019151:Peptide Library; D015203:Reproducibility of Results; D013678:Technology, Pharmaceutical / Q000379:methods
chemical_list: D000906:Antibodies; D019151:Peptide Library