- Research on gold nanoparticles (AuNPs) is being continued from the previous year. AuNPs were synthesized with bovine serum albumin (BSA) in H2O at various mole ratios of Au/BSA and their presence in solution was analyzed with UV-vis spectroscopy and atomic absorption spectroscopy. Different methods of synthesizing AuNPs are being studied, including using adenosine deaminase (ADA) or lysozyme rather than BSA.
- A Bradford assay was used to determine the amount of ADA obtained from protein expression. ADA was also mutated with K110A forward and reverse primers and underwent protein expression using E. coli.
- AuNPs were synthesized with lysozyme at various mole ratios of Au/lysozyme and their presence in solution was analyzed with UV-vis and atomic absorption spectroscopy.
- AuNPs were also synthesized with ADA and horseradish peroxidase (HRP) using a similar protocol and also analyzed with UV-vis and AAS.
Protein Activity Assays
- Two activity assays of HRP were optimized and the activity of HRP was calculated from the data. The first activity assay used aminoantipyrine (AAP) as the substrate and the reaction was monitored with UV-vis spectroscopy. The second activity assay monitored the fluorescence of luminol in the presence of HRP.
- An activity assay of ADA by measuring the absorbance of adenosine or inosine in solution is being optimized. The ultimate goal is to conduct the assay on ADA that has been conjugated to AuNPs to determine whether the activity of ADA is hindered by AuNPs.