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Journal Article
Methods
. 2013 Sep; 65(1):57-67.
doi: 10.1016/j.ymeth.2013.09.003.

Retrocyte Display® technology: generation and screening of a high diversity cellular antibody library

Ekaterina Breous-Nystrom 1,  Kornelia Schultze 1,  Marco Meier 1,  Lukas Flueck 1,  Christina Holzer 1,  Melanie Boll 1,  Volker Seibert 1,  Andrea Schuster 1,  Milan Blanusa 1,  Verena Schaefer 1,  Ulf Grawunder 2,  Luis Martin-Parras 1,  Marc A van Dijk 3 
Affiliations
  • PMID: 24036249
  •     12 citations

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).

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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