1. Retrocyte Display® technology: generation and screening of a high diversity cellular antibody library.
- Author
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Breous-Nystrom E, Schultze K, Meier M, Flueck L, Holzer C, Boll M, Seibert V, Schuster A, Blanusa M, Schaefer V, Grawunder U, Martin-Parras L, and van Dijk MA
- Subjects
- Animals, Antibodies, Monoclonal genetics, B-Lymphocytes metabolism, Cryopreservation, Drug Evaluation, Preclinical, Flow Cytometry, Genetic Variation, Genetic Vectors, HEK293 Cells, Humans, Immunoglobulin Heavy Chains biosynthesis, Immunoglobulin Heavy Chains genetics, Immunoglobulin Light Chains biosynthesis, Immunoglobulin Light Chains genetics, Immunomagnetic Separation, Peptide Library, Protein Binding, Antibodies, Monoclonal biosynthesis, Retroviridae genetics
- 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)., (Copyright © 2013 Elsevier Inc. All rights reserved.)
- Published
- 2014
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