title: Determination of glycan motifs using serial lectin affinity chromatography.
abstract: Serial lectin affinity chromatography is a convenient technique for characterizing glycan motifs (terminal glycan structures) of glycoproteins or released glycans. When these glycoconjugates are applied serially or in parallel to lectin-immobilized columns, information regarding the glycan motifs can be obtained. We demonstrate lectin affinity chromatographic methods for determining O-linked glycan structures of MUC1 purified from a breast cancer cell line, YMB-S, N-linked glycan structures of serum prostate-specific antigen from prostate cancer, and serum alkaline phosphatases from choriocarcinoma. These lectin-fractionated samples are analyzed quantitatively by measuring radioactivity, antigen contents are analyzed using enzyme-linked immunosorbent assay, and enzymatic activities are assessed.
authors.affiliation: Department of Histology and Cell Biology, School of Medicine, Yokohama City University, 3-9, Fukuura, Kanazawa-ku, 236-0004, Yokohama, Japan, kyamashita34@gmail.com.
authors.forename: Katsuko
authors.initials: K
authors.lastname: Yamashita
authors.forename: Takashi
authors.initials: T
authors.lastname: Ohkura
mesh_terms: D045744:Cell Line, Tumor; D002846:Chromatography, Affinity / Q000379:methods*; D006026:Glycoside Hydrolases / Q000378:metabolism; D006801:Humans; D055767:Immobilized Proteins / Q000737:chemistry / Q000378:metabolism; D037102:Lectins / Q000737:chemistry / Q000378:metabolism*; D008297:Male; D018396:Mucin-1 / Q000097:blood / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D011134:Polysaccharides / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D011470:Prostatic Hyperplasia / Q000097:blood; D011471:Prostatic Neoplasms / Q000097:blood
chemical_list: D055767:Immobilized Proteins; D037102:Lectins; C060192:MUC1 protein, human; D018396:Mucin-1; D011134:Polysaccharides; D006026:Glycoside Hydrolases