title: Influence of immobilization protocol on the structure and function of surface bound proteins.
abstract: A new coupling strategy for biomacromolecules with (3-mercaptopropyl)trimethoxysilane (3MPTMS) and 11-(triethoxysilyl)undecanal (TESU) on gold surfaces is. This immobilization protocol was utilized for the enzyme horseradish peroxidase (HRP). To study the reactions and resulting structures, PM-IRRAS measurements were performed. PM-IRRAS shows there is structure preservation of the HRP when the new coupling strategy is used in contrast to non-specific adsorption on gold. The biological activity of adsorbed and immobilized HRP was measured by the enzyme catalyzed oxidation of 3,5,3',5'-tetramethylbenzidine. Covalent immobilization of HRP on TESU film compared to physisorption of HRP shows higher enzyme activity on gold surfaces, confirming the structural preservation detected by PM-IRRAS.
authors: Fraunhofer Institute for Manufacturing Technology and Advanced Materials, Wiener Strasse 12, Bremen 28359, Germany.
authors: Alexej
authors: A
authors: Kreider
authors: Fraunhofer Institute for Manufacturing Technology and Advanced Materials, Wiener Strasse 12, Bremen 28359, Germany.
authors: Stephan
authors: S
authors: Sell
authors: Fraunhofer Institute for Manufacturing Technology and Advanced Materials, Wiener Strasse 12, Bremen 28359, Germany.
authors: Thomas
authors: T
authors: Kowalik
authors: Fraunhofer Institute for Manufacturing Technology and Advanced Materials, Wiener Strasse 12, Bremen 28359, Germany. Electronic address: andreas.hartwig@ifam.fraunhofer.de.
authors: Andreas
authors: A
authors: Hartwig
authors: Fraunhofer Institute for Manufacturing Technology and Advanced Materials, Wiener Strasse 12, Bremen 28359, Germany.
authors: Ingo
authors: I
authors: Grunwald
mesh_terms: D000327:Adsorption; D004789:Enzyme Activation; D004800:Enzymes, Immobilized / Q000737:chemistry / Q000378:metabolism*; D006046:Gold / Q000737:chemistry / Q000378:metabolism; D006735:Horseradish Peroxidase / Q000737:chemistry / Q000378:metabolism*; D008958:Models, Molecular; D015394:Molecular Structure; D017646:Organosilicon Compounds; D012821:Silanes / Q000737:chemistry / Q000378:metabolism; D013055:Spectrophotometry, Infrared; D013499:Surface Properties
chemical_list: D004800:Enzymes, Immobilized; D017646:Organosilicon Compounds; D012821:Silanes; D006046:Gold; C102833:(3-mercaptopropyl)trimethoxysilane; D006735:Horseradish Peroxidase
keywords: Bioactivity; Biofunctionalization; Covalent immobilization; Horseradish peroxidase; PM-IRRAS