Sugar-binding proteins from fish: selection of high affinity "lambodies" that recognize biomedically relevant glycans.

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TitleSugar-binding proteins from fish: selection of high affinity "lambodies" that recognize biomedically relevant glycans.
Publication TypeJournal Article
Year of Publication2013
AuthorsHong, X, Ma, MZ, Gildersleeve, JC, Chowdhury, S, Barchi, JJ, Mariuzza, RA, Murphy, MB, Mao, L, Pancer, Z
JournalACS Chem Biol
Volume8
Issue1
Pagination152-60
Date Published2013 Jan 18
ISSN1554-8937
KeywordsAnimals, Antibodies, Monoclonal, Antigens, Neoplasm, Blotting, Western, Carbohydrate Sequence, Cell Line, Tumor, Flow Cytometry, Humans, Lampreys, Lung Neoplasms, Microarray Analysis, Molecular Sequence Data, Polysaccharides, Protein Binding, Reference Standards
Abstract

Glycan-binding proteins are important for a wide variety of basic research and clinical applications, but proteins with high affinity and selectivity for carbohydrates are difficult to obtain. Here we describe a facile and cost-effective strategy to generate monoclonal lamprey antibodies, called lambodies, that target glycan determinants. We screened a library of yeast surface-displayed (YSD) lamprey variable lymphocyte receptors (VLR) for clones that can selectively bind various biomedically important glycotopes. These glycoconjugates included tumor-associated carbohydrate antigens (Tn and TFα), Lewis antigens (LeA and LeX), N-glycolylneuraminic acid, targets of broadly neutralizing HIV antibodies (poly-Man9 and the HIV gp120), and the glycoproteins asialo-ovine submaxillary mucin (aOSM) and asialo-human glycophorin A (aGPA). We isolated clones that bind each of these targets in a glycan-dependent manner and with very strong binding constants, for example, 6.2 nM for Man9 and 44.7 nM for gp120, determined by surface plasmon resonance (SPR). One particular lambody, VLRB.aGPA.23, was shown by glycan array analysis to be selective for the blood group H type 3 trisaccharide (BG-H3, Fucα1-2Galβ1-3GalNAcα), aGPA, and TFα (Galβ1-3GalNAcα), with affinity constants of 0.2, 1, and 8 nM, respectively. In human tissue microarrays this lambody selectively detected cancer-associated carbohydrate antigens in 14 different types of cancers. It stained 27% of non-small cell lung cancer (NSCLC) samples in a pattern that correlated with poor patient survival. Lambodies with exquisite affinity and selectivity for glycans may find myriad uses in glycobiology and biomedical research.

DOI10.1021/cb300399s
Alternate JournalACS Chem. Biol.
PubMed ID23030719
PubMed Central IDPMC3756686
Grant ListAI083892 / AI / NIAID NIH HHS / United States
CA126818 / CA / NCI NIH HHS / United States
CA136635 / CA / NCI NIH HHS / United States
GM098791 / GM / NIGMS NIH HHS / United States
GM62116 / GM / NIGMS NIH HHS / United States
R01 CA126818 / CA / NCI NIH HHS / United States
R01 CA136635 / CA / NCI NIH HHS / United States
R21 AI083892 / AI / NIAID NIH HHS / United States