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

Jeong Heon Lee; Yangsun Kim; Mi Young Ha; Eun Kyu Lee; Jaebum Choo

doi:10.1016/j.jasms.2005.04.005

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

J Am Soc Mass Spectrom. 2005 Sep;16(9):1456-1460. doi: 10.1016/j.jasms.2005.04.005.

Authors

Jeong Heon Lee¹, Yangsun Kim², Mi Young Ha³, Eun Kyu Lee³, Jaebum Choo⁴

Affiliations

¹ Proteonik Research Laboratory, Ansan, South Korea.
² Proteonik Research Laboratory, Ansan, South Korea. yangskim@microbiochip.com.
³ Microbiochip Center, Hanyang University, Ansan, South Korea.
⁴ Department of Applied Chemistry, Hanyang University, Ansan, South Korea.

PMID: 16023361
DOI: 10.1016/j.jasms.2005.04.005

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.

Publication types

Evaluation Study
Research Support, Non-U.S. Gov't

MeSH terms

Absorption
Boronic Acids / chemistry*
Coated Materials, Biocompatible / chemistry*
Glycoproteins / analysis
Glycoproteins / chemistry*
Immunomagnetic Separation / methods*
Microspheres
Protein Binding
Specimen Handling / methods
Spectrometry, Mass, Electrospray Ionization / methods*
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / methods*

Substances

Boronic Acids
Coated Materials, Biocompatible
Glycoproteins
3-aminobenzeneboronic acid