TY - JOUR
T1 - Structural analyses of O-glycan sugar chains on IgA1 hinge region using SELDI-TOFMS with various lectins
AU - Takahashi, Kazuo
AU - Hiki, Yoshiyuki
AU - Odani, Hiroko
AU - Shimozato, Sachiko
AU - Iwase, Hitoo
AU - Sugiyama, Satoshi
AU - Usuda, Nobuteru
N1 - Funding Information:
The authors are grateful to Prof. A. Ishii and Dr. R. Kaneko (Department of Legal Medicine, Fujita Health University, School of Medicine) for valuable suggestions and to Ms. I. Matsumoto, Ms. K. Inoue, and Ms. M. Itoh for their superb technical assistance. This study was in part supported by a grant-in-aid for the 21st Century Center of Excellence Program of Fujita Health University from the Ministry of Education, Culture, Sports, Sciences, and Technology of Japan, the Ministry of Education and Science (No. 16590840), and Aichi Kidney Foundation.
PY - 2006/11/24
Y1 - 2006/11/24
N2 - The aim of the study was to develop a simple and precise method for identifying glycosylation of the IgA hinge region using surface-enhanced laser desorption/ionization (SELDI)-TOFMS with a lectin-coupled ProteinChip array. Serum IgA was isolated using an anti-IgA antibody column. Following reduction, alkylation, and trypsin digestion, the IgA fragments were applied on the ProteinChip coupled with jacalin, peanut agglutinin (PNA), or Vilsa villosa lectin (VVL). The SELDI-TOFMS peaks corresponding to the fragments containing IgA1 hinge glycopeptides trapped by each lectin were compared. The jacalin-, PNA-, and VVL-immobilized ProteinChips detected 13, 4, and 2 peaks, respectively. One major peak was confirmed as a glycopeptide by MS/MS analysis. These results suggest that a lectin-immobilized ProteinChip assay can be used to simplify the procedures for the analyses of the O-glycans in IgA1 hinge. This method potentially makes it possible to identify a disease-specific glycoform by selecting the appropriate ligand-coupled ProteinChip array.
AB - The aim of the study was to develop a simple and precise method for identifying glycosylation of the IgA hinge region using surface-enhanced laser desorption/ionization (SELDI)-TOFMS with a lectin-coupled ProteinChip array. Serum IgA was isolated using an anti-IgA antibody column. Following reduction, alkylation, and trypsin digestion, the IgA fragments were applied on the ProteinChip coupled with jacalin, peanut agglutinin (PNA), or Vilsa villosa lectin (VVL). The SELDI-TOFMS peaks corresponding to the fragments containing IgA1 hinge glycopeptides trapped by each lectin were compared. The jacalin-, PNA-, and VVL-immobilized ProteinChips detected 13, 4, and 2 peaks, respectively. One major peak was confirmed as a glycopeptide by MS/MS analysis. These results suggest that a lectin-immobilized ProteinChip assay can be used to simplify the procedures for the analyses of the O-glycans in IgA1 hinge. This method potentially makes it possible to identify a disease-specific glycoform by selecting the appropriate ligand-coupled ProteinChip array.
UR - https://www.scopus.com/pages/publications/33749598630
UR - https://www.scopus.com/pages/publications/33749598630#tab=citedBy
U2 - 10.1016/j.bbrc.2006.09.075
DO - 10.1016/j.bbrc.2006.09.075
M3 - Article
C2 - 17022936
AN - SCOPUS:33749598630
SN - 0006-291X
VL - 350
SP - 580
EP - 587
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
IS - 3
ER -