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J Immunol Methods. 2012 Dec 14;386(1-2):43-9. doi: 10.1016/j.jim.2012.08.016. Epub 2012 Sep 4.

Purification and identification of cell surface antigens using lamprey monoclonal antibodies.

Author information

1
Department of Pathology and Laboratory Medicine, Emory University School of Medicine, 1462 Clifton Rd NE, Room 403C, Atlanta, GA 30322, USA.

Abstract

Variable lymphocyte receptor (VLR) B antibodies of the evolutionary distant sea lamprey are structurally distinct from conventional mammalian antibodies. The different protein architecture and large evolutionary distance of jawless vertebrates suggest that VLR antibodies may represent promising tools for biomarker discovery. Here we report the generation of panels of monoclonal VLR antibodies from lamprey larvae immunized with human T cells and the use of a recombinant monoclonal VLR antibody for antigen purification and mass spectrometric identification. We demonstrate that despite predicted low affinity of individual VLR antigen binding units to the antigen, the high avidity resulting from decameric assembly of secreted VLR antibodies allows for efficient antigen capture and subsequent identification by mass spectometry. We show that VLR antibodies detect their antigens with high specificity and can be used in various standard laboratory application techniques. The lamprey antibodies are novel reagents that can complement conventional monoclonal antibodies in multiple scientific research disciplines.

PMID:
22964555
PMCID:
PMC3756502
DOI:
10.1016/j.jim.2012.08.016
[Indexed for MEDLINE]
Free PMC Article

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