Journal Article
. 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.

Transpo-mAb display: Transposition-mediated B cell display and functional screening of full-length IgG antibody libraries.
Lorenz Waldmeier, Ina Hellmann, +5 authors, Roger R Beerli.
MAbs, 2016 Mar 18; 8(4). PMID: 26986818    Free PMC article.
Toxicological and pharmacological assessment of AGEN1884, a novel human IgG1 anti-CTLA-4 antibody.
Randi B Gombos, Ana Gonzalez, +24 authors, Nicholas S Wilson.
PLoS One, 2018 Apr 05; 13(4). PMID: 29617360    Free PMC article.
Novel Antibody Drug Conjugates Targeting Tumor-Associated Receptor Tyrosine Kinase ROR2 by Functional Screening of Fully Human Antibody Libraries Using Transpo-mAb Display on Progenitor B Cells.
Ina Hellmann, Lorenz Waldmeier, +4 authors, Roger R Beerli.
Front Immunol, 2018 Nov 20; 9. PMID: 30450096    Free PMC article.
Antibodies to watch in 2020.
Hélène Kaplon, Mrinalini Muralidharan, Zita Schneider, Janice M Reichert.
MAbs, 2019 Dec 19; 12(1). PMID: 31847708    Free PMC article.
Highly Cited. Review.
A comprehensive search of functional sequence space using large mammalian display libraries created by gene editing.
Kothai Parthiban, Rajika L Perera, +8 authors, John McCafferty.
MAbs, 2019 May 21; 11(5). PMID: 31107136    Free PMC article.
Enhancers Improve the AID-Induced Hypermutation in Episomal Vector for Antibody Affinity Maturation in Mammalian Cell Display.
Chuan Chen, Jie Wang, +4 authors, Haiying Hang.
Antibodies (Basel), 2019 Sep 24; 7(4). PMID: 31544892    Free PMC article.
Beyond affinity: selection of antibody variants with optimal biophysical properties and reduced immunogenicity from mammalian display libraries.
Michael R Dyson, Edward Masters, +11 authors, John McCafferty.
MAbs, 2020 Oct 27; 12(1). PMID: 33103593    Free PMC article.
Beyond antibody engineering: directed evolution of alternative binding scaffolds and enzymes using yeast surface display.
Doreen Könning, Harald Kolmar.
Microb Cell Fact, 2018 Feb 28; 17(1). PMID: 29482656    Free PMC article.
Review.
Streamlined human antibody generation and optimization by exploiting designed immunoglobulin loci in a B cell line.
Hidetaka Seo, Hitomi Masuda, +15 authors, Yukoh Nakazaki.
Cell Mol Immunol, 2020 May 28; 18(6). PMID: 32457406    Free PMC article.
Development of a novel mammalian display system for selection of antibodies against membrane proteins.
Nathan Robertson, Nancy Lopez-Anton, +10 authors, Ryan Cawood.
J Biol Chem, 2020 Nov 01; 295(52). PMID: 33127646    Free PMC article.
Balstilimab and other immunotherapy for recurrent and metastatic cervical cancer.
Chinmoy K Bose.
Med Oncol, 2022 Jan 30; 39(4). PMID: 35092506
Review.
Next-Generation Molecular Discovery: From Bottom-Up In Vivo and In Vitro Approaches to In Silico Top-Down Approaches for Therapeutics Neogenesis.
Sophie E Kenny, Fiach Antaw, +3 authors, Matt Trau.
Life (Basel), 2022 Mar 26; 12(3). PMID: 35330114    Free PMC article.
Review.