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Updated: Dec 30, 2025

Covalent Immobilization of Proteins for the Single Molecule Force Spectroscopy
Published on: August 20, 2018
Immobilization of Enzymes on Hetero-Functional Supports: Physical Adsorption Plus Additional Covalent Immobilization
Lara Trobo-Maseda1, Alejandro H Orrego1, María Romero-Fernández1
1Department of Biocatalysis, Institute of Catalysis and Petrochemistry (ICP) CSIC, Campus UAM, Madrid, Spain.
Abstract:
The immobilization of proteins on heterofunctional amino-epoxy and amino-glyoxyl supports is described in this chapter. Immobilization on both supports is performed through a two-step mechanism: in the first step, the enzyme is physically adsorbed to the support, and in the second step, the intramolecular covalent attachment between the adsorbed enzyme and the support is promoted. On the one hand, amino-epoxy supports present a ratio between amino and epoxy groups of 1:1 to allow the rapid adsorption of the enzyme and promote a strong multipoint covalent linkage. On the other hand, amino-glyoxyl supports contain the highest concentration of glyoxyl groups capable of reacting covalently with primary amino groups on the enzyme surface to promote increased rigidification. The intensity of the covalent enzyme-support interaction can be modulated by modifying the ratio between glyoxyl and amino groups of the support. These heterofunctional supports are able to immobilize and rigidify proteins through different orientations, leading to biocatalysts with different enzyme properties (activity, stability, and selectivity).
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