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Retrocyte Display® technology: generation and screening of a high diversity cellular antibody library.
- Source :
-
Methods (San Diego, Calif.) [Methods] 2014 Jan 01; Vol. 65 (1), pp. 57-67. Date of Electronic Publication: 2013 Sep 12. - Publication Year :
- 2014
-
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).<br /> (Copyright © 2013 Elsevier Inc. All rights reserved.)
- 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
Subjects
Details
- Language :
- English
- ISSN :
- 1095-9130
- Volume :
- 65
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- Methods (San Diego, Calif.)
- Publication Type :
- Academic Journal
- Accession number :
- 24036249
- Full Text :
- https://doi.org/10.1016/j.ymeth.2013.09.003