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Updated: May 3, 2026

Covalent Immobilization of Proteins for the Single Molecule Force Spectroscopy
Published on: August 20, 2018
Influence of immobilization protocol on the structure and function of surface bound proteins
Alexej Kreider1, Stephan Sell1, Thomas Kowalik1
1Fraunhofer Institute for Manufacturing Technology and Advanced Materials, Wiener Strasse 12, Bremen 28359, Germany.
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.
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