Quantitative LC-MS/MS method for nivolumab in human serum using IgG purification and immobilized tryptic digestion

A simple and rapid LC-MS/MS method for quantification of nivolumab in human serum using immobilized trypsin coupled to IgG purification.

Immobilization of Trypsin from Porcine Pancreas onto Chitosan Nonwoven by Covalent Bonding

The present study deals with the potential application of chitosan nonwoven for biomedical textiles based on enzyme immobilization. For this, chitosan nonwoven was first cross-linked with glutaraldehyde to introduce aldehyde groups at optimal conditions. To immobilize the enzyme trypsin onto glutaraldehyde-pre-activated chitosan nonwoven, several parameters such as pH, enzyme concentration, and reaction times were investigated. In addition, the pH, thermal stability, storage stability, and reusability of immobilized trypsin were examined. We found that the optimal immobilization conditions for trypsin were pH 8.5, enzyme concentration of 8% (owf), and treatment time of 30 min. Trypsin was immobilized at 25 °C efficiently. The immobilized trypsin showed lower pH stability and better thermal stability than free trypsin. The immobilized trypsin showed 50% of its initial activity after being used 15 times and 80% of that after 20 days of storage at 4 °C. SEM analysis also confirmed that trypsin was immobilized on chitosan nonwoven.