Engineered PlyCB as Novel Implant Coating for Osseointegration
Abstract
During the performance period, we successfully constructed five RGD or cyclic-RGD containing mutants to PlyCB. Four of them were to the N-terminus and one was to the external loop between helicies 1 and 2 of PlyCB. All five mutants were expressed, purified, and analyzed to confirm retention of wild-type properties. These tests included analytical gel filtration/dynamic light scattering (to confirm self assembly of PlyCB into an octameric ring) and thermal denaturation studies by circular dichroism (to confirm proper protein folding and thermalstability properties). We found that addition of RGD or cyclic RGD did not affect any of the wild-type biochemical or biophysical properties when added to the N-terminus of PlyCB. Three N-terminal mutants were chemically crosslinked to a fluorophore for visualization by fluorescent microscopy in later aims. Next, we tested the binding of mutants to hydroxyapatite (HA) under various salt, pH, phosphate, and temperature conditions. Under all conditions, the mutants strongly and completely bound HA.
Document Details
- Document Type
- Technical Report
- Publication Date
- Sep 01, 2011
- Accession Number
- ADA554576
Entities
People
- Daniel C Nelson
Organizations
- University of Maryland