Abstract

We present a technology to screen millions of B cells for natively paired human antibody repertoires. Libraries of natively paired, variable region heavy and light (VH:VL) amplicons are expressed in a yeast display platform that is optimized for human Fab surface expression. Using our method we identify HIV-1 broadly neutralizing antibodies (bNAbs) from an HIV-1 slow progressor and high-affinity neutralizing antibodies against Ebola virus glycoprotein and influenza hemagglutinin.

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GenBank/EMBL/DDBJ

Sequence Read Archive

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Acknowledgements

We gratefully acknowledge B. Hartman for assistance with figures, N. Doria-Rose, G. Ippolito, and M. Kanekiyo for advice and guidance, S. Lucas, and K. Zhou for help with experiments, S. Darko for assistance with data processing, and E. Shusta and S. Harrison for kindly sharing reagents and research tools. This work was funded in part by the intramural research program of the Vaccine Research Center, NIAID, NIH, NIH grant DP5OD023118-01 to B.J.D., NIH grant 5R21CA191239-01 to A.D.E., Leidos Biomedical Research Inc. contract 15X219, DTRA contract HDTRA1-12-C-0105, and NIH grant 1R56AI106006 to G.G.

Author information

Author notes

    • Bo Wang
    •  & Brandon J DeKosky

    These authors contributed equally to this work.

Affiliations

  1. Department of Chemical Engineering, The University of Texas at Austin, Austin, Texas, USA.

    • Bo Wang
    • , Jiwon Lee
    • , Jonathan R McDaniel
    • , George Delidakis
    • , William N Voss
    •  & George Georgiou
  2. Vaccine Research Center, National Institute of Allergy and Infectious Diseases, Bethesda, Maryland, USA.

    • Brandon J DeKosky
    • , Morgan R Timm
    • , Erica Normandin
    • , John Misasi
    • , Rui Kong
    • , Kendra E Leigh
    • , Thomas Niezold
    • , Chang W Choi
    • , Elise G Viox
    • , Alberto Cagigi
    • , Aurélie Ploquin
    • , Kwanyee Leung
    • , Eun Sung Yang
    • , Wing-Pui Kong
    • , David R Ambrozak
    • , Amy R Henry
    • , Farida Laboune
    • , Julie E Ledgerwood
    • , Barney S Graham
    • , Daniel C Douek
    • , Nancy J Sullivan
    •  & John R Mascola
  3. Department of Chemical & Petroleum Engineering, The University of Kansas, Lawrence, Kansas, USA.

    • Brandon J DeKosky
  4. Department of Pharmaceutical Chemistry, The University of Kansas, Lawrence, Kansas, USA.

    • Brandon J DeKosky
    •  & Ahmed Fahad
  5. Laboratory of Molecular Medicine, Boston Children′s Hospital, Harvard Medical School, Boston, Massachusetts, USA.

    • Aaron G Schmidt
  6. Duke Human Vaccine Institute, Duke University Medical School, Durham, North Carolina, USA.

    • M Anthony Moody
  7. Department of Pediatrics, Duke University Medical School, Durham, North Carolina, USA.

    • M Anthony Moody
  8. National Institute of Allergy and Infectious Diseases, Bethesda, Maryland, USA.

    • Mark Connors
  9. Center for Systems and Synthetic Biology, The University of Texas at Austin, Austin, Texas, USA.

    • Andrew D Ellington
    •  & George Georgiou
  10. Department of Molecular Biosciences, The University of Texas at Austin, Austin, Texas, USA.

    • Andrew D Ellington
    •  & George Georgiou
  11. Department of Bioengineering, The University of Texas at Austin, Austin, Texas, USA.

    • George Georgiou

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Contributions

B.W., B.J.D., J.R. Mascola, and G.G. conceived the study and designed the experiments. B.W. and B.J.D. conducted the experiments with help from M.R.T., J.L., E.N., J.M., R.K., J.R. McDaniel, G.D., K.E.L., T.N., C.W.C., E.G.V., A.F., A.C., A.P., K.L., E.S.Y., W.-P.K., W.N.V., A.G.S., M.A.M., D.R.A., A.R.H., F.L., J.E.L., B.S.G., M.C., and D.C.D. N.J.S, A.D.E., J.R. Mascola, and G.G. supervised the study. B.W., B.J.D., J.R. Mascola, and G.G. wrote the manuscript with input from all authors.

Competing interests

Competing financial interests statement G.G., B.J.D., and A.D.E. declare competing financial interests in the form of a patent filed by the University of Texas at Austin.

Corresponding authors

Correspondence to John R Mascola or George Georgiou.

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DOI

https://doi.org/10.1038/nbt.4052