title: Isolation and characterization of monoclonal antibodies specific for chondroitin sulfate E.
abstract: Chondroitin sulfate E (CSE) is a polysaccharide containing mainly disaccharide units of D-glucuronic acid (GlcA) and 4,6-O-disulfated N-acetyl-D-galactosamine (GalNAc) residues (E-unit) in the amount of ∼ 60%. CSE is involved in many biological and pathological processes. In this study, we established new monoclonal antibodies, termed E-12C and E-18H, by using CSE that contained more than 70% of E-units as an immunogen. These antibodies recognized CSE but not other CSs isomers or dermatan sulfate (DS). We evaluated the reactivities of the antibodies to 6-O-sulfated CSA (6S-CSA) and DS (6S-DS) that possessed ∼ 60% of GalNAc (4S, 6S) moieties in their structures. Neither of the antibodies reacted with 6S-DS. The antibodies strictly distinguished the structural difference of GlcA and L-iduronic acid in the polysaccharide. Binding affinities of the antibodies were determined by a surface plasmon resonance assay using CSE and 6S-CSA. The binding affinities were strongly associated with the molecular weight of CSE and the E-unit content of 6S-CSA. Moreover, we demonstrated that the antibodies are applicable to histochemical analysis. In conclusion, the new anti-CSE monoclonal antibodies specifically recognize the E-unit of CSE. The antibodies will become useful tools for the investigation of the biological and pathological significance of CSE.
authors.affiliation: Department of Biochemistry, School of Pharmaceutical Science, University of Shizuoka, 52-1 Yada, Suruga-ku, Shizuoka-shi, Shizuoka 422-8526, Japan Central Research Laboratories, Seikagaku Corporation, 3-1253 Tateno, Higashiyamato-shi, Tokyo 207-0021, Japan.
authors.forename: Ippei
authors.initials: I
authors.lastname: Watanabe
authors.affiliation: Department of Biochemistry, School of Pharmaceutical Science, University of Shizuoka, 52-1 Yada, Suruga-ku, Shizuoka-shi, Shizuoka 422-8526, Japan.
authors.forename: Tomoya
authors.initials: T
authors.lastname: Hikita
authors.affiliation: Department of Biochemistry, School of Pharmaceutical Science, University of Shizuoka, 52-1 Yada, Suruga-ku, Shizuoka-shi, Shizuoka 422-8526, Japan.
authors.forename: Haruka
authors.initials: H
authors.lastname: Mizuno
authors.affiliation: Department of Biochemistry, School of Pharmaceutical Science, University of Shizuoka, 52-1 Yada, Suruga-ku, Shizuoka-shi, Shizuoka 422-8526, Japan.
authors.forename: Risa
authors.initials: R
authors.lastname: Sekita
authors.affiliation: Department of Biochemistry, School of Pharmaceutical Science, University of Shizuoka, 52-1 Yada, Suruga-ku, Shizuoka-shi, Shizuoka 422-8526, Japan.
authors.forename: Akira
authors.initials: A
authors.lastname: Minami
authors.affiliation: Department of Biochemistry, School of Pharmaceutical Science, University of Shizuoka, 52-1 Yada, Suruga-ku, Shizuoka-shi, Shizuoka 422-8526, Japan.
authors.forename: Ami
authors.initials: A
authors.lastname: Ishii
authors.affiliation: Central Research Laboratories, Seikagaku Corporation, 3-1253 Tateno, Higashiyamato-shi, Tokyo 207-0021, Japan.
authors.forename: Yuka
authors.initials: Y
authors.lastname: Minamisawa
authors.affiliation: Central Research Laboratories, Seikagaku Corporation, 3-1253 Tateno, Higashiyamato-shi, Tokyo 207-0021, Japan.
authors.forename: Kiyoshi
authors.initials: K
authors.lastname: Suzuki
authors.affiliation: Central Research Laboratories, Seikagaku Corporation, 3-1253 Tateno, Higashiyamato-shi, Tokyo 207-0021, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Maeda
authors.affiliation: Department of Food and Nutrition, Junior College Division, University of Aizu, 1-1 Aza-Kadota Yahata, Ikki-machi, Aizuwakamatsu-shi, Fukushima 965-8570, Japan.
authors.forename: Kazuya I P J
authors.initials: KI
authors.lastname: Hidari
authors.affiliation: Department of Biochemistry, School of Pharmaceutical Science, University of Shizuoka, 52-1 Yada, Suruga-ku, Shizuoka-shi, Shizuoka 422-8526, Japan suzukit@u-shizuoka-ken.ac.jp.
authors.forename: Takashi
authors.initials: T
authors.lastname: Suzuki
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology*; D000915:Antibody Affinity; D000918:Antibody Specificity; D045744:Cell Line, Tumor; D002809:Chondroitin Sulfates / Q000276:immunology*; D006801:Humans; D051379:Mice; D051381:Rats
chemical_list: D000911:Antibodies, Monoclonal; D002809:Chondroitin Sulfates
keywords: binding affinity; chondroitin sulfate E; glycosaminoglycan; monoclonal antibody; surface plasmon resonance