Activity of enzyme immobilized on silanized Co‐Cr‐Mo
- 1 August 1995
- journal article
- research article
- Published by Wiley in Journal of Biomedical Materials Research
- Vol. 29 (8), 951-957
- https://doi.org/10.1002/jbm.820290806
Abstract
The surface of an orthopedic biomaterial was modified by the covalent immobilization of biomolecules. Derivatization of Co-Cr-Mo samples with organic and aqueous solutions of γ-aminopropyltriethoxysilane (APS) resulted in a concentration-dependent number of reactive NH2 groups on the surface available for coupling to protein. The enzyme trypsin was used as a model biomolecule to investigate the effect of immobilization on proteolytic activity. Trypsin was coupled to the silanized samples by formation of Schiff's base linkages via glutaraldehyde. The nature of the interaction between trypsin and biomaterial was then probed by treatment with concentrated guanidine hydrochloride (Gu-HCl) and urea. Residual activity (following treatment with chaotropic agents) of trypsin immobilized on silanized Co-Cr-Mo was dependent both on the nature of the silane solution and on the type of chaotropic agent. Organic silanization with APS required a miniumum density of approximately 49 NH2 per nm2 of nominal surface area (>0.021 M APS) for residual activity of immobilized trypsin. For aqueous silanization, approximately 5.4 NH2/nm2 (0.51 M APS) resulted in maximal residual trypsin activity. Treatment with GuHCl removed more trypsin activity from Co-Cr-Mo samples silanized with organic solutions of APS than did treatment with urea. On the contrary, with aqueous silanization the samples possessed greater residual activity followig treatment with GuHCl than following urea. Compared to simple adsorption with protein onto Co-Cr-Mo, both methods of silanization with APS resulted in superior residual immobilized enzyme activity. © 1995 John Wiley & Sons, Inc.Keywords
This publication has 22 references indexed in Scilit:
- Structure, solubility and bond strength of thin calcium phosphate coatings produced by ion beam sputter depositionBiomaterials, 1992
- Human endothelial cell interactions with surface‐coupled adhesion peptides on a nonadhesive glass substrate and two polymeric biomaterialsJournal of Biomedical Materials Research, 1991
- Plasma‐sprayed coatings of tetracalciumphosphate, hydroxyl‐apatite, and α‐TCP on titanium alloy: An interface studyJournal of Biomedical Materials Research, 1991
- Modulation of cell adhesion by modification of titanium surfaces with covalently attached self‐assembled monolayersJournal of Biomedical Materials Research, 1990
- Total Hip and Total Knee ReplacementNew England Journal of Medicine, 1990
- A nonantigenic covalent streptokinase-polyethylene glycol complex with plasminogen activator function.JCI Insight, 1985
- Improved nonporous magnetic supports for immobilized enzymesBiotechnology & Bioengineering, 1979
- A Family of Protein-Cutting ProteinsScientific American, 1974
- Determination of the amino acid sequence of porcine trypsin by sequenator analysisBiochemistry, 1973
- Nonpolar Group Participation in the Denaturation of Proteins by Urea and Guanidinium Salts. Model Compound StudiesJournal of the American Chemical Society, 1964