Use of novel fluorescent probes to monitor Protein interactions with medical devices
|Co-investigator(s):||A.C. Sullivan and Karin HING|
This programme of work involved development of a method to facilitate the competitive evaluation of protein adsorption to dense and porous apatites via covalent labelling of the species of interest with a fluorescent probe so that it may be monitored independently of other protein species in the local environment. Fluoresceinthiureidoaminocaproic acid (FTCA) and a sulforhodamine derivative (SR101), were identified as suitable probes for coupling to proteins as both parent fluorophores are known to have no adverse effect on cells or proteins, and have proven stability in several relevant media. This work has resulted in accurate quantification of the degree of sensitivity of dynamic protein exchange to both the environment (media composition and temperature) and device surface physico-chemistry (surface morphology and chemistry), and highlighted that in situations where protein conformation is radically altered during interation with medical devices, traditional ELISA type assays can be unrelable.