Preparation of reusable enzyme strip for determination of serum triglyceride

Abstract

Lipase, glycerol kinase (GK), glycerol-3-phosphate oxidase and peroxidase were co-immobilized onto alkylamine glass beads affixed on a plastic strip with a conjugation yield of 80.0 mg/g support and 57.41% retention of specific activity. The enzyme showed an increase in optimum pH, incubation temperature, energy of activation and time of incubation, but decrease in <i style="">K</i>_mand <i style="">V</i>_max. A method for determination of serum triglycerides was developed, employing strip bound enzymes. The minimum detection limit of the method was 0.61 mM/L. The recovery of added triglycerides was 82.2%. Within and between assay coefficient of variation were < 3.1 and < 14.2 respectively. A good correlation (<i style="">r</i>=0.973) was found between triglycerides value obtained by commercial enzymatic colourimetric method and present method. The strip provided 100 reuses of co-immobilized enzymes with ease.