Fabrication and characterization of L-asparaginase entrapped polymeric nanoparticles for asparagine biosensor construction

Abstract

The present work reports the fabrication of novel and highly stable L-asparaginase (L-ASP) loaded gelatin alginate nanoparticles (GANp) for the construction of an easily reproducible, sensitive, and selective fiber optic asparagine biosensor. GANp containing L-ASP of 396 nm size with 79.7% entrapment efficiency was achieved via ionic gelation method. The physicochemical properties of fabricated GANp were characterized by Scanning Electron Microscopy (SEM) and Fourier Transform Infrared Spectroscopy (FTIR). The fabricated L-ASP entrapped GANp were stable for 50 days with 75% of residual activity. Immobilized L-ASP displayed improved stability against pH and temperature with higher affinity (Km3.83 mM) in comparison to free L-ASP. Furthermore, the application of L-ASP loaded GANp (biocomponent) in biosensor construction led to the evolution of highly sensitive fiber optic asparagine biosensor with 0.05 × 10-10 M detection limit within 2 min of response time. The proposed biosensor can be potentially applied for L-asparagine detection and quantification in the leukemia diagnosis and treatment.