| Literature DB >> 24036249 |
Ekaterina Breous-Nystrom1, Kornelia Schultze1, Marco Meier1, Lukas Flueck1, Christina Holzer1, Melanie Boll1, Volker Seibert1, Andrea Schuster1, Milan Blanusa1, Verena Schaefer1, Ulf Grawunder2, Luis Martin-Parras1, Marc A van Dijk3.
Abstract
Over the last nearly three decades in vitro display technologies have played an important role in the discovery and optimization of antibodies and other proteins for therapeutic applications. Here we describe the use of retroviral expression technology for the display of full-length IgG on B lineage cells in vitro with a hallmark of a tight and stable genotype to phenotype coupling. We describe the creation of a high-diversity (>1.0E09 different heavy- and light-chain combinations) cell displayed fully human antibody library from healthy donor-derived heavy- and light-chain gene libraries, and demonstrate the recovery of high affinity target-specific antibodies from this library by staining of cells with a labeled target antigen and their magnetic- and flow cytometry-based cell sorting. The present technology represents a further evolution in the discovery of full-length, fully human antibodies using mammalian display, and is termed Retrocyte Display® (Retroviral B lymphocyte Display).Entities:
Keywords: A-MuLV; Abelson murine leukemia virus; B cell receptor; BCR; BiP; CDR; DR6; FACS; FCS; Flow cytometry; Fully human antibody; IRES; MACS; Mammalian cell surface antibody display; Retrocyte Display®; Retroviral B lymphocyte Display; Retroviral transduction; TNF-α; VH; Vκ; binding immunoglobulin protein; complementarity determining region; death receptor 6; fetal calf serum; fluorescence activated cell sorting; heavy chain; internal ribosome entry site; kappa light-chain; magnetic activated cell sorting; tumor necrosis factor-alpha
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Year: 2013 PMID： 24036249 DOI： 10.1016/j.ymeth.2013.09.003
Source DB: PubMed Journal: Methods ISSN： 1046-2023 Impact factor: 3.608