Facile Conjugation of Biomolecules onto Surfaces via Mussel Adhesive Protein Inspired Coatings.

Abstract

Immobilization of biomolecules onto surfaces is important in many of the biological and physical sciences, including cell and molecular biology, analytical chemistry, and in applied and interdisciplinary fields such as medical diagnostics, tissue engineering, and bioprocess engineering.[1-4] Strategies for biomolecule immobilization onto surfaces generally exploit either noncovalent or covalent reactions. Noncovalent methods allow reversible immobilization of biomolecules under specific conditions, and include physical adsorption and affinity immobilization. Some widely adapted examples are (strep)avidin-biotin, nitriloacetic acid (NTA)-histidine, and DNA-DNA interactions.[5-8] In contrast, covalent immobilization of molecules onto surfaces typically relies on conjugation reactions between ‘active’ functional groups, such as N-hydroxysuccinimide (NHS)[9] or maleimide,[10] and companion target moieties, such as amines and sulfhydryls. For reactions involving biomolecules performed in aqueous solvents, susceptibility of NHS, maleimide, and other activating groups to hydrolysis during storage and reaction can lead to low efficiency of surface bioconjugation.[11,12] In this study, we report a facile two-step aqueous approach to immobilization of biomolecules onto surfaces. The approach involves simple dip-coating of a biomimetic polymer thin film onto a substrate, followed by conjugation of biomolecules to the biomimetic polymer film. The method exploits the latent reactivity of the biomimetic polymer thin film towards nucleophiles, is unaffected by water, and allows for discrimination between nucleophiles on the basis of pKa.