Premium
Modification of aluminum chips for LDI mass spectrometry of proteins
Author(s) -
Shmanai Vadim,
Gontarev Sergey,
Frey Simone K.,
Schweigert Florian J.
Publication year - 2007
Publication title -
journal of mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.475
H-Index - 121
eISSN - 1096-9888
pISSN - 1076-5174
DOI - 10.1002/jms.1259
Subject(s) - chemistry , agarose , chromatography , boronic acid , matrix assisted laser desorption/ionization , mass spectrometry , polymer , affinity chromatography , covalent bond , glycation , desorption , adsorption , combinatorial chemistry , organic chemistry , biochemistry , enzyme , receptor
Matrix‐assisted laser desorption ionization time‐of‐flight mass spectroscopy (MALDI TOFMS) combined with affinity chromatography on immobilized phenylboronic acid agarose gels was used for selective enrichment and detection of specifically modified proteins such as glycated proteins in complex biological samples. Physicochemical grafting of hydrophilic polymers on aluminum surface was developed to reduce nonspecific protein sorption and to create a proper support layer for a three‐dimensional affinity hydrogel. Grafted agarose allowed the fixation of three‐dimensional agarose hydrogel on the chip surface. Both pinched polymers and hydrogels were effectively derivatized. 3‐Aminophenylboronic acid ( m PBA) was covalently immobilized as an affinity ligand to achieve specific binding of glycated plasma proteins. Alternatively, the affinity sorbent was immersed into the hydrogel to increase binding capacity. MALDI TOFMS was used to evaluate binding efficiency and molecular mass changes of human serum albumin due to glycation. Glycated proteins were captured directly on the chip with high selectivity and efficacy, and low nonspecific binding. Thus they could easily be characterized by MALDI TOFMS. Copyright © 2007 John Wiley & Sons, Ltd.