Generation of Lamprey Monoclonal Antibodies (Lampribodies) Using the Phage Display System

• Type of Material: Article
• Authors: Khan M. A. Hassan, University of California Merced; John D. Hansen, Western Fisheries Research Center; Brantley Herrin, Emory University; Chris T. Amemiya, University of California Merced
• Language: English
• Date: 2019-12-01
• Publisher: MDPI
• Publication Version: Final Published Version
• Copyright Statement: © 2019 by the authors.
• License: Creative Commons Attribution 4.0 International
• Final Published Version (URL): http://dx.doi.org/10.3390/biom9120868
• Title of Journal or Parent Work: Biomolecules
• Volume: 9
• Issue: 12
• Start Page: 868
• End Page: 868
• Grant/Funding Information: This work was funded, in part, by the Washington Research Foundation and discretionary funds from the Benaroya Research Institute and University of California-Merced.; Initial parts of this work were carried out at the Benaroya Research Institute, Seattle, Washington.; Research at Emory University was funded by NIH grant R01AI072435 (to Cooper).
• Keywords: variable lymphocyte receptors; phage display system; lampribody; library screening
• Research Categories: Biology, Molecular; Biology, Cell

Abstract

The variable lymphocyte receptors (VLRs) consist of leucine rich repeats (LRRs) and comprise the humoral antibodies produced by lampreys and hagfishes. The diversity of the molecules is generated by stepwise genomic rearrangements of LRR cassettes dispersed throughout the VLRB locus. Previously, target-specific monovalent VLRB antibodies were isolated from sea lamprey larvae after immunization with model antigens. Further, the cloned VLR cDNAs from activated lamprey leukocytes were transfected into human cell lines or yeast to select best binders. Here, we expand on the overall utility of the VLRB technology by introducing it into a filamentous phage display system. We first tested the efficacy of isolating phage into which known VLRB molecules were cloned after a series of dilutions. These experiments showed that targeted VLRB clones could easily be recovered even after extensive dilutions (1 to 109). We further utilized the system to isolate target-specific “lampribodies” from phage display libraries from immunized animals and observed an amplification of binders with relative high affinities by competitive binding. The lampribodies can be individually purified and ostensibly utilized for applications for which conventional monoclonal antibodies are employed.