A quantitative study of the enzymatic activity of horseradish peroxidase at a planar poly(acrylic acid) brush.
The enzymatic activity of horseradish peroxidase (HRP) has been quantified at a planar poly(acrylic acid) (PAA) brush. A PAA brush is known to exhibit an unusual protein binding affinity, since proteins adsorb at low ionic strength only. At elevated ionic strengths of a few 100 mM proteins desorb, and the PAA brush becomes largely protein resistant. In this study, HRP was used to catalyze the oxidation of Amplex Red to resorufin by hydrogen peroxide. The fluorescence of resorufin was recorded using a microplate reader. As compared to a bare silica surface, the enzymatic activity of HRP is higher by more than one order of magnitude at a PAA brush. This increase results from a higher degree of adsorption and a reasonable preservation of the HRP activity. Upon adsorption at a PAA brush from a 20 mU/mL (0.1 microg/mL) solution, the molecular enzymatic activity of HRP is reduced to about 11%. However, when a HRP molecule is desorbed from a PAA brush by increasing the ionic strength, a gain of the molecular enzymatic activity by only 52% can be observed. Overall, this study illustrates the potential applicability of a PAA brush as a biocompatible material coating.
Reichhart
Christian
C
Technische Universität Dortmund, Fakultät Chemie, D-44221 Dortmund, Germany.
Czeslik
Claus
C
D000180:Acrylic Resins / Q000378:metabolism*; D000327:Adsorption / Q000187:drug effects; D057075:Enzyme Assays / Q000379:methods*; D005453:Fluorescence; D006735:Horseradish Peroxidase / Q000378:metabolism*; D010078:Oxazines / Q000378:metabolism; D010084:Oxidation-Reduction / Q000187:drug effects; D012822:Silicon Dioxide / Q000378:metabolism; D012965:Sodium Chloride / Q000494:pharmacology; D012996:Solutions; D013997:Time Factors; D014867:Water / Q000378:metabolism
D000180:Acrylic Resins; D010078:Oxazines; D012996:Solutions; D014867:Water; D012965:Sodium Chloride; C006903:carbopol 940; C014180:resorufin; D012822:Silicon Dioxide; D006735:Horseradish Peroxidase