Lentivirus Display: Stable expression of human antibodies on the surface of human cells and virus particles

Ran Taube, Quan Zhu, Chen Xu, Felipe Diaz-Griffero, Jianhua Sui, Erick Kamau, Markryan Dwyer, Daniel Aird, Wayne A. Marasco

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Background: Isolation of human antibodies using current display technologies can be limited by constraints on protein expression, folding and post-translational modifications. Here we describe a discovery platform that utilizes self-inactivating (SIN) lentiviral vectors for the surface display of high-affinity single-chain variable region (scFv) antibody fragments on human cells and lentivirus particles. Methodalogy/Principal Findings: Bivalent scFvFc human antibodies were fused in frame with different transmembrane (TM) anchoring moieties to allow efficient high-level expression on human cells and the optimal TM was identified. The addition of an eight amino acid HIV-1 gp41 envelope incorporation motif further increased scFvFc expression on human cells and incorporation into lentiviral particles. Both antibody-displaying human cells and virus particles bound antigen specifically. Sulfation of CDR tyrosine residues, a property recently shown to broaden antibody binding affinity and antigen recognition was also demonstrated. High level scFvFc expression and stable integration was achieved in human cells following transduction with IRES containing bicistronic SIN lentivectors encoding ZsGreen when scFvFc fusion proteins were expressed from the first cassette. Up to 106-fold enrichment of antibody expressing cells was achieved with one round of antigen coupled magnetic bead pre-selection followed by FACS sorting. Finally, the scFvFc displaying human cells could be used directly in functional biological screens with remarkable sensitivity. Conclusions/Significance: This antibody display platform will complement existing technologies by virtue of providing properties unique to lentiviruses and antibody expression in human cells, which, in turn, may aid the discovery of novel therapeutic human mAbs.

Original languageEnglish (US)
Article numbere3181
JournalPLoS One
Volume3
Issue number9
DOIs
StatePublished - Sep 11 2008
Externally publishedYes

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Lentivirus
Viruses
virion
Virion
Display devices
Cells
antibodies
Antibodies
cells
Antigens
antigens
Single-Chain Antibodies
Immunoglobulin Fragments
Sorting
Technology
Tyrosine
Antibody Affinity
Proteins
Fusion reactions
Protein Folding

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Lentivirus Display : Stable expression of human antibodies on the surface of human cells and virus particles. / Taube, Ran; Zhu, Quan; Xu, Chen; Diaz-Griffero, Felipe; Sui, Jianhua; Kamau, Erick; Dwyer, Markryan; Aird, Daniel; Marasco, Wayne A.

In: PLoS One, Vol. 3, No. 9, e3181, 11.09.2008.

Research output: Contribution to journalArticle

Taube, Ran ; Zhu, Quan ; Xu, Chen ; Diaz-Griffero, Felipe ; Sui, Jianhua ; Kamau, Erick ; Dwyer, Markryan ; Aird, Daniel ; Marasco, Wayne A. / Lentivirus Display : Stable expression of human antibodies on the surface of human cells and virus particles. In: PLoS One. 2008 ; Vol. 3, No. 9.
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