Abstract
High-quality reagents to study and detect glycans with high specificity for research and clinical applications are severely lacking. Here, we structurally and functionally characterize several variable lymphocyte receptor (VLR)-based antibodies from lampreys immunized with O erythrocytes that specifically recognize the blood group H-trisaccharide type II antigen. Glycan microarray analysis and biophysical data reveal that these VLRs exhibit greater specificity for H-trisaccharide compared with the plant lectin UEA-1, which is widely used in blood typing. Among these antibodies, O13 exhibits superior specificity for H-trisaccharide, the basis for which is revealed by comparative analysis of high-resolution VLR:glycan crystal structures. Using a structure-guided approach, we designed an O13 mutant with further enhanced specificity for H-trisaccharide. These insights into glycan recognition by VLRs suggest that lampreys can produce highly specific glycan antibodies, and are a valuable resource for the production of next-generation glycan reagents for biological and biomedical research and as diagnostics and therapeutics.
Keywords:
X-ray crystallography; glycans; glycobiology; glycomics; immunology; leucine-rich repeat; structural biology; variable lymphocyte receptor.
Copyright © 2017 Elsevier Ltd. All rights reserved.
Publication types
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Research Support, U.S. Gov't, Non-P.H.S.
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Research Support, N.I.H., Extramural
MeSH terms
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Amino Acid Sequence
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Animals
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Antibodies, Monoclonal / biosynthesis
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Antibodies, Monoclonal / chemistry*
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Antibodies, Monoclonal / isolation & purification
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Antibody Specificity
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Binding Sites
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Blood Group Antigens / analysis*
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Blood Group Antigens / immunology
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Blood Grouping and Crossmatching / methods
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Crystallography, X-Ray
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Erythrocytes / chemistry
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Erythrocytes / immunology
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Humans
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Immunization
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Lampreys / immunology*
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Models, Molecular
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Plant Lectins / chemistry
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Plant Lectins / immunology
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Polysaccharides / chemistry*
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Polysaccharides / immunology
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Polysaccharides / metabolism
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Protein Binding
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Protein Conformation
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Protein Conformation, alpha-Helical
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Protein Conformation, beta-Strand
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Protein Interaction Domains and Motifs
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Receptors, Antigen, T-Cell / chemistry*
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Receptors, Antigen, T-Cell / genetics
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Receptors, Antigen, T-Cell / immunology
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Recombinant Proteins / chemistry
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Recombinant Proteins / genetics
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Recombinant Proteins / metabolism
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Sequence Alignment
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Sequence Homology, Amino Acid
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Trisaccharides / chemistry*
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Trisaccharides / immunology
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Trisaccharides / metabolism
Substances
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Antibodies, Monoclonal
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Blood Group Antigens
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Plant Lectins
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Polysaccharides
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Receptors, Antigen, T-Cell
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Recombinant Proteins
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Trisaccharides
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Ulex europaeus lectins