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Contributed by Richard A. Lerner, May 25, 2018 (sent for review April 19, 2018; reviewed by Richard A. Houghten and Sachdev S. Sidhu)
Significance
We constructed a library of miniecosystems that can translate the information from antibody phage display directly into signals of biological function, thereby allowing for rapid selection of antibodies with the function of interest. Compared with the conventional phage display platform that can only isolate antibodies based on their binding affinity toward antigens, our new method bypasses the step of affinity-based selection, and the selection is based purely on the activity of antibodies in a biological system without concern for their relative affinity for antigens. This new method bridges the gap, which has existed for almost three decades, between affinity- and activity-based antibody selection for phage display of combinatorial antibody libraries, thus advancing antibody drug discovery.
Abstract
We describe a method for the rapid selection of functional antibodies. The method depends on the cocultivation of Escherichia coli that produce phage with target eukaryotic cells in very small volumes. The antibodies on phage induce selectable phenotypes in the target cells, and the nature of the antibody is determined by gene sequencing of the phage genome. To select functional antibodies from the diverse antibody repertoire, we devised a selection platform that contains millions of picoliter-sized droplet ecosystems. In each miniecosystem, the bacteria produce phage displaying unique members of the antibody repertoire. These phage interact only with eukaryotic cells in the same miniecosystem, making phage available directly for activity-based antibody selection in biological systems.
Footnotes
- ↵1To whom correspondence should be addressed. Email: rlerner{at}scripps.edu.
Author contributions: T.Z. and R.A.L. designed research; T.Z., Z.Y., P.T., B.S., and L.D. performed research; J.X. contributed new reagents/analytic tools; T.Z., Z.Y., P.T., B.S., L.D., P.W., and R.A.L. analyzed data; and T.Z. and R.A.L. wrote the paper.
Reviewers: R.A.H. Torrey Pines Institute for Molecular Studies; and S.S.S., University of Toronto.
The authors declare no conflict of interest.
This article contains supporting information online at www.pnas.org/lookup/suppl/doi:10.1073/pnas.1806718115/-/DCSupplemental.
Published under the PNAS license.
Antibody selection using clonal cocultivation of Escherichia coli and eukaryotic cells in miniecosystems
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