Yeast Surface Display of Lamprey Variable Lymphocyte Receptors

Protocol
First Online:
Part of the Methods in Molecular Biology book series (MIMB, volume 748)

Abstract

The variable lymphocyte receptors (VLRs) of lamprey and hagfish comprise leucine-rich repeat modules, instead of the immunoglobulin-like domain building blocks of antibodies and T-cell receptors in jawed vertebrates. Both types of vertebrate-rearranging antigen receptors are similarly diverse, with repertoires that can potentially exceed 1014 unique receptors. In order to characterize antigen-binding properties of the VLRs, we developed a high-throughput yeast surface display platform for the isolation of monoclonal VLRs. We have isolated VLRs that specifically bind hen egg lysozyme, β-galactosidase, cholera toxin subunit B, R-phycoerythrin, and the blood group trisaccharides A and B, with binding affinities in the mid-nanomolar to mid-picomolar range. VLRs may, thus, be excellent single-chain alternatives to Ig-based antibodies for biotechnology applications.

Key words

Lamprey Variable lymphocyte receptors Recombinant antibodies Yeast surface display 
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Notes

Acknowledgments

The authors would like to thank Dr. K. Dane Wittrup and Ms. S. Annie Gai from Massachusetts Institute of Technology for introducing us to the fascinating world of yeast surface display (YSD). This work was supported by NSF grant MCB-0614672, NIH grant AI083892, and Intercenter Collaboration Grant, UMBI.

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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  1. 1.Center of Marine BiotechnologyUniversity of Maryland Biotechnology InstituteBaltimoreUSA

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