CE detection of N-(3-dimethylaminopropyl)-N-carbodiimide/N-hydroxysuccinimide-coupled proteins after homo- and hetero-crosslinking reactions.
Protein-protein conjugates formed by carbodiimide crosslinking reactions have been analyzed for the first time using CE. Lysozyme and BSA were chosen as model proteins to study the efficacy of N-(3-dimethylaminopropyl)-N-ethylcarbodiimide and N-hydroxysuccinimide as crosslinkers. Detection of the molecular mass increase was checked by SDS-PAGE. Commercially available, PVA-coated capillaries showed appropriate selection, while phospho-deactivated and dynamic PVA-coated capillaries did not give suitable resolution. CE was found to be an efficient tool to characterize homo- (lysozyme-lysozyme) and hetero- (lysozyme-BSA) protein coupling by suitable variations of electrophoretic mobilities.
Muskotál
Adél
A
Institute of Engineering Materials and Design, Faculty of Mechanical Engineering, University of Maribor, Maribor, Slovenia.
Kokol
Vanja
V
D003432:Cross-Linking Reagents / Q000737:chemistry*; D019075:Electrophoresis, Capillary / Q000379:methods*; D005022:Ethyldimethylaminopropyl Carbodiimide / Q000737:chemistry*; D009113:Muramidase / Q000737:chemistry; D012710:Serum Albumin, Bovine / Q000737:chemistry; D013388:Succinimides / Q000737:chemistry*
D003432:Cross-Linking Reagents; D013388:Succinimides; D012710:Serum Albumin, Bovine; D009113:Muramidase; C001426:N-hydroxysuccinimide; D005022:Ethyldimethylaminopropyl Carbodiimide