Purification and identification of cell surface antigens using lamprey monoclonal antibodies

• Type of Material: Article
• Authors: Cuiling Yu, Emory University; Shabab Ali, University of Toronto; Jonathan St-Germain, Hospital for Sick Children; Yanling Liu, University of Toronto; Xuecong Yu, University of Toronto; David L Jaye, Emory University; Michael F. Moran, Hospital for Sick Children; Max Dale Cooper, Emory University; Götz R.A. Ehrhardt, University of Toronto
• Language: English
• Date: 2012-12-14
• Publisher: Elsevier
• Publication Version: Author Accepted Manuscript (After Peer Review)
• Copyright Statement: © 2012 Elsevier B.V.
• License: Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International
• Final Published Version (URL): http://dx.doi.org/10.1016/j.jim.2012.08.016
• Title of Journal or Parent Work: Journal of Immunological Methods
• ISSN: 0022-1759
• Volume: 386
• Issue: 1-2
• Start Page: 43
• End Page: 49
• Grant/Funding Information: This study was supported in part by Canadian Cancer Society grant 2012-701054 to G. Ehrhardt, NIH grant 5U19AI096187-02 to G. Ehrhardt and M. Cooper and NIH grant 2R01AI072435-07 to M. Cooper.
• Keywords: VLR antibody; Science & Technology; Mass spectrometry; Biochemistry & Molecular Biology; Antigen identification; DIVERSIFICATION; VARIABLE LYMPHOCYTE RECEPTOR; Life Sciences & Biomedicine; Evolution; SUBEPITHELIAL B-CELLS; Monoclonal; Immunology; EVOLUTION; Biochemical Research Methods; RECOGNITION
• Research Categories: Health Sciences, Pathology; Biology, Genetics

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. © 2012 Elsevier B.V.