title: Characterization of anti-heparan sulfate phage display antibodies AO4B08 and HS4E4.
abstract: Heparan sulfates (HS) are linear carbohydrate chains, covalently attached to proteins, that occur on essentially all cell surfaces and in extracellular matrices. HS chains show extensive structural heterogeneity and are functionally important during embryogenesis and in homeostasis due to their interactions with various proteins. Phage display antibodies have been developed to probe HS structures, assess the availability of protein-binding sites, and monitor structural changes during development and disease. Here we have characterized two such antibodies, AO4B08 and HS4E4, previously noted for partly differential tissue staining. AO4B08 recognized both HS and heparin, and was found to interact with an ubiquitouys, N-, 2-O-, and 6-O-sulfated saccharide motif, including an internal 2-O-sulfate group. HS4E4 turned out to preferentially recognize low-sulfated HS motifs containing iduronic acid, and N-sulfated as well as N-acetylated glucosamine residues. Contrary to AO4B08, HS4E4 did not bind highly O-sulfated structures such as found in heparin.
authors.affiliation: Department of Medical Biochemistry and Microbiology, University of Uppsala, SE-751 23 Sweden.
authors.forename: Sindhulakshmi
authors.initials: S
authors.lastname: Kurup
authors.forename: Tessa J M
authors.initials: TJ
authors.lastname: Wijnhoven
authors.forename: Guido J
authors.initials: GJ
authors.lastname: Jenniskens
authors.forename: Koji
authors.initials: K
authors.lastname: Kimata
authors.forename: Hiroko
authors.initials: H
authors.lastname: Habuchi
authors.forename: Jin-Ping
authors.initials: JP
authors.lastname: Li
authors.forename: Ulf
authors.initials: U
authors.lastname: Lindahl
authors.forename: Toin H
authors.initials: TH
authors.lastname: van Kuppevelt
authors.forename: Dorothe
authors.initials: D
authors.lastname: Spillmann
mesh_terms: D020816:Amino Acid Motifs; D000818:Animals; D000906:Antibodies / Q000737:chemistry*; D001665:Binding Sites; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D000939:Epitopes / Q000737:chemistry; D005944:Glucosamine / Q000737:chemistry; D006497:Heparan Sulfate / Q000737:chemistry* / Q000276:immunology; D007067:Iduronic Acid / Q000737:chemistry; D019151:Peptide Library; D011134:Polysaccharides / Q000737:chemistry; D011485:Protein Binding; D017434:Protein Structure, Tertiary
chemical_list: D000906:Antibodies; D000939:Epitopes; D005944:Glucosamine; D006497:Heparan Sulfate; D007067:Iduronic Acid; D019151:Peptide Library; D011134:Polysaccharides