Synthesis and characterization of cysteine functionalized silver nanoparticles for biomolecule immobilization.
A facile method for the aqueous phase synthesis of cysteine-functionalized silver nanoparticles by potato extract has been reported in the present work. These functionalized nanoparticles were then used for the covalent immobilization of a biomolecule, alkaline phosphatase, on its surface through carbodiimide coupling. Different reaction parameters such as cysteine concentration, reducing agent concentration, temperature, pH and reaction time were varied during the nanoparticles' formation, and their effects on plasmon resonance were studied using Ultraviolet-visible spectroscopy. Fourier transform infrared spectroscopy was used to confirm the surface modification of silver nanoparticles by cysteine and the particle size analysis was done using particle size analyzer, which showed the average nanoparticles' size of 61 nm for bare silver nanoparticles and 201 nm for the enzyme-immobilized nanoparticles. The synthesized nanoparticles were found to be highly efficient for the covalent immobilization of alkaline phosphatase on its surface and retained 67% of its initial enzyme activity (9.44 U/mg), with 75% binding efficiency. The shelf life of the enzyme-nanoparticle bioconjugates was found to be 60 days, with a 12% loss in the initial enzyme activity. With a simple synthesis strategy, high immobilization efficiency and enhanced stability, these enzyme-coated nanoparticles have the potential for further integration into the biosensor technology.
Department of Biotechnology, National Institute of Technology, Raipur, Raipur, 492001, Chhattisgarh, India, contactlataupadhyay@gmail.com.
Lata Sheo Bachan
LS
Upadhyay
Nishant
N
Verma
D000469:Alkaline Phosphatase; D015374:Biosensing Techniques / Q000379:methods; D003544:Cysteic Acid / Q000737:chemistry*; D004795:Enzyme Stability; D004800:Enzymes, Immobilized; D006863:Hydrogen-Ion Concentration; D053768:Metal Nanoparticles / Q000737:chemistry* / Q000648:ultrastructure; D008855:Microscopy, Electron, Scanning; D036103:Nanotechnology; D010316:Particle Size; D012834:Silver / Q000737:chemistry*; D013056:Spectrophotometry, Ultraviolet; D017550:Spectroscopy, Fourier Transform Infrared; D013696:Temperature
D004800:Enzymes, Immobilized; D012834:Silver; D003544:Cysteic Acid; D000469:Alkaline Phosphatase