Efficient method to optimize antibodies using avian leukosis virus display and eukaryotic cells

Changming Yu, Gennett M. Pike, Tommy A. Rinkoski, Cristina Correia, Scott H Kaufmann, Mark J Federspiel, Robin A. Weiss

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Antibody-based therapeutics have now had success in the clinic. The affinity and specificity of the antibody for the target ligand determines the specificity of therapeutic delivery and off-target side effects. The discovery and optimization of high-affinity antibodies to important therapeutic targets could be significantly improved by the availability of a robust, eukaryotic display technology comparable to phage display that would overcome the protein translation limitations of microorganisms. The use of eukaryotic cells would improve the diversity of the displayed antibodies that can be screened and optimized as well as more seamlessly transition into a large-scale mammalian expression system for clinical production. In this study, we demonstrate that the replication and polypeptide display characteristics of a eukaryotic retrovirus, avian leukosis virus (ALV), offers a robust, eukaryotic version of bacteriophage display. The binding affinity of a model single-chain Fv antibody was optimized by using ALV display, improving affinity >2,000-fold, from micromolar to picomolar levels. We believe ALV display provides an extension to antibody display on microorganisms and offers virus and cell display platforms in a eukaryotic expression system. ALV display should enable an improvement in the diversity of properly processed and functional antibody variants that can be screened and affinity-optimized to improve promising antibody candidates.

Original languageEnglish (US)
Pages (from-to)9860-9865
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume112
Issue number32
DOIs
StatePublished - Aug 11 2015

Fingerprint

Avian Leukosis Virus
Eukaryotic Cells
Antibody Affinity
Antibodies
Bacteriophages
Antibody Diversity
Single-Chain Antibodies
Antibody Specificity
Protein Biosynthesis
Retroviridae
Therapeutics
Ligands
Viruses
Technology
Peptides

Keywords

  • Antibody binding affinity
  • Antibody engineering
  • Avian leukosis virus
  • Avian leukosis virus polypeptide display
  • Protein display technology

ASJC Scopus subject areas

  • General

Cite this

Efficient method to optimize antibodies using avian leukosis virus display and eukaryotic cells. / Yu, Changming; Pike, Gennett M.; Rinkoski, Tommy A.; Correia, Cristina; Kaufmann, Scott H; Federspiel, Mark J; Weiss, Robin A.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 112, No. 32, 11.08.2015, p. 9860-9865.

Research output: Contribution to journalArticle

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