Development of a sensitive method for selection of affinity ligand for trypsin using quartz crystal microbalance sensor

摘要

In this work, a new methodology is developed for selection of affinity ligands towards the enzyme "trypsin" using quartz crystals microbalance (QCM) technique. To achieve this goal, the surface amination of gold plated QCM crystals was achieved in 13.56 MHz plasma polymerization system by using ethylenediamine. Three different ligands (i.e., 4-aminobenzamidine, 4-amino-benzoic acid, and phenylalanine) were immobilized on the aminated QCM crystals surface via glutaraldhyde coupling. All three ligand immobilized QCM crystals were characterized and compared under different experimental conditions. It was observed that the benzamidine ligand showed higher affinity to trypsin with a dissociation constant on the order of 1.76 × 10~9 M, which is within the range of 10~(-4)-10~(-8) M for affinity ligands. Thus, its selectivity was suitable for purification of trypsin from biological fluids.