(95l) Examination of the Biological Stability of Myoglobin from Horse Skeletal Muscle and Hemoglobin I from Lucina Pectinata in Monomeric and Solvent Enviroments

This project proposes to investigate the immobilization of Hemoglobin I from Lucina pectinata (HbI)with the entrapment technique for Hydrogen Sulfide biosensor application. This technique provides the means to create dense networks thus significantly decreasing the amount of protein that could be released, creating a network that may be employed as a biosensor. However the biological activity of the protein could be severely hindered by the monomer and solvent composition. Since HbI is not commercially available and must be isolated and purified, myoglobin from horse skeletal muscle (Mb) was employed to develop all the necessary experimental protocols. Anionic hydrogels composed of methacrylic acid (MAA), cross-linked with poly(ethylene glycol) dimethacrylate (PEGDMA) (n=200, n=1000), were synthesized in presence of the protein by free radical solution polymerization. The stability of the protein was examined in the pre-polymeric solution since acids and ethanol can denature the protein by disrupting the hydrophilic and hydrophobic interaction that make up the stable core of the protein. Results indicated that a solvent composition of 30% of ethanol in deionized water do not affect the protein activity. Others components such as methacrylic acid and the cross-linker will be analyzed. The activity of the protein was determined using UV spectroscopy. Release studies will be conducted to evaluate the amount of protein permanently immobilized inside the hydrogels and to evaluate the diffusion mechanism during release.