Protein immobilization on polyvinylphenol via tyrosine oxidation of proteins catalyzed by horseradish peroxidase

Abstract

AbstractSurface functionalization of polymeric materials holds great potential in biological and medical fields with the aim toward realizing desired biological reactions (e.g., cell adhesion, and immune response), especially for proteins which have physiological functions. The objective was to examine characteristics of a unique and facile protein immobilization on a phenol‐containing polymeric material utilizing horseradish peroxidase (HRP) and tyrosine residues of target proteins as catalyst and substrates, respectively. Cy3‐labeled streptavidin exhibited the predicted immobilization behavior since the reaction effectively proceeded in the presence of HRP‐H2O2 and on the surface of polyvinylphenol. This procedure was applicable to other representative proteins, that is, bovine serum albumin, immunoglobulin G (IgG), and HRP as substrates. However, IgG exhibited anomalous reaction behavior under the present reaction condition probably due to its rich tyrosine content. Undesired immobilization of HRP was minimized by addition of large amounts of a competitive substrate. The biotin‐binding affinity of streptavidin‐immobilized surface was confirmed to maintain activity even after the immobilization procedure.