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Immobilization of aminophenylboronic acid on magnetic beads for the direct determination of glycoproteins by matrix assisted laser desorption ionization mass spectroscopy

Title
Immobilization of aminophenylboronic acid on magnetic beads for the direct determination of glycoproteins by matrix assisted laser desorption ionization mass spectroscopy
Author
이은규
Keywords
MALDI; GlcNAc; Magnetic Bead; Direct Determination; Confocal Laser Microscopy
Issue Date
2005-09
Publisher
SPRINGER
Citation
JOURNAL OF THE AMERICAN SOCIETY FOR MASS SPECTROMETRY, v. 16, No. 9, Page. 1456-1460
Abstract
Aminophenylboronic acid (APBA) has been immobilized on magnetic beads for the direct determination of glycoprotein by matrix assisted laser desorption/ionizaton time of flight mass spectrometry (MALDI-TOF-MS). An APBA layer was formed on the surface of carboxylic acid terminated magnetic beads by coupling with carbodiimide and subsequently reacted with an N-hydroxysuccinimide moiety. The immobilized APBA was identified by MALDI-TOF-MS without a matrix. Glycoproteins, such as HbA1c, fibrinogen, or RNase B were separated and desalted using APBA magnetic beads by simply washing the magnetic beads and then separating them by external magnet. Proteins can be identified by direct determination of proteins on beads on MALDI plate and confirmed again by peptide mass finger printing after digestion of proteins on magnetic beads by trypsin. Fluorescence image with a FITC tagging protein using confocal laser microscopy showed the difference of immobilization efficiency between glycoproteins and nonglycoproteins. The methods developed within this work allow the simple treatment and enrichment of glycoproteins as well as direct determination of proteins on beads by MALDI-TOF-MS.
URI
https://link.springer.com/article/10.1016/j.jasms.2005.04.005https://repository.hanyang.ac.kr/handle/20.500.11754/111186
ISSN
1044-0305; 1879-1123
DOI
10.1016/j.jasms.2005.04.005
Appears in Collections:
COLLEGE OF ENGINEERING SCIENCES[E](공학대학) > BIONANO ENGINEERING(생명나노공학과) > Articles
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