Application of phenylboronic acid modified hydrogel affinity chips for high-throughput mass spectrometric analysis of glycated proteins.
The complexity of the human plasma proteome is greatly increased by post-translational modifications. Besides physiological modifications, pathological conditions such as diabetes are responsible for adding to this complexity by producing advanced glycation endproducts (AGEs). When searching for specific biomarkers it is a prerequisite to reduce this complexity prior to analysis. To do this, agarose hydrogel was used to create a high-capacity affinity layer on the modified aluminum surface of MALDI (matrix-assisted laser desorption/ionization) targets. 3-Aminophenylboronic acid was immobilized via cyanogen bromide activation as a ligand for affinity sorption of glycated proteins, followed by their direct detection by MALDI. High protein capacity of prepared MALDI chips, efficient separation and low non-specific protein binding were demonstrated. The results show that phenylboronic acid modified hydrogels are very suitable for creating affinity surfaces for the high-throughput analysis of AGEs.
Gontarev
Sergey
S
Bioanalyt GmbH, Arthur-Scheunert-Allee 114-116, 14558 Nuthetal, Germany.
Shmanai
Vadim
V
Frey
Simone K
SK
Kvach
Maksim
M
Schweigert
Florian J
FJ
D000535:Aluminum / Q000737:chemistry; D001897:Boronic Acids / Q000032:analysis*; D003488:Cyanogen Bromide; D005453:Fluorescence; D017127:Glycation End Products, Advanced / Q000032:analysis*; D006023:Glycoproteins / Q000032:analysis*; D020100:Hydrogels / Q000032:analysis*; D007202:Indicators and Reagents; D008024:Ligands; D013058:Mass Spectrometry / Q000379:methods*; D040081:Protein Array Analysis; D040901:Proteomics / Q000379:methods*; D012685:Sepharose / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013056:Spectrophotometry, Ultraviolet
D001897:Boronic Acids; D017127:Glycation End Products, Advanced; D006023:Glycoproteins; D020100:Hydrogels; D007202:Indicators and Reagents; D008024:Ligands; D012685:Sepharose; D000535:Aluminum; C010686:benzeneboronic acid; D003488:Cyanogen Bromide