Determination of serum glucose using co-immobilized glucose oxidase and peroxidase onto arylamine glass beads affixed on a plastic strip.
Glucose oxidase (GOD) from Aspergillus niger and horseradish peroxidase (POD) were co-immobilized onto arylamine glass beads affixed on a plastic strip with a conjugation yield of 28.2 mg/g and 43% retention of their initial specific activity. The coimmobilized enzymes showed maximum activity at pH 7.5 when incubated at 37 degrees C for 15 min. A simple, specific and sensitive method for discrete analysis of the serum glucose was developed employing this strip. The minimum detection limit of the method was 5 mg/dl. Within and between assay coefficient of variations for the serum were <5.6% and <10.6% (n = 6) respondely. A good correlation (r = 0.943) was found between the glucose values obtained by the enzyme colorimetric method employing free GOD and POD and the present method. The strip lost 50% of its initial activity after its 150 regular uses for a period of one month, when stored in reaction buffer at 4 degrees C. The method is cost-effective than the enzymic colorimetric method, as the enzyme strip is reusable.
Tank
Neelima
N
Biochemistry Research Laboratory, Dept of Bio Science, M D University, Rohtak, Haryana, India.
Suman
Pundir
C S
CS
D000328:Adult; D000588:Amines; D001234:Aspergillus niger / Q000201:enzymology*; D001786:Blood Glucose / Q000378:metabolism*; D003920:Diabetes Mellitus / Q000097:blood*; D004800:Enzymes, Immobilized / Q000378:metabolism*; D005260:Female; D005898:Glass; D005949:Glucose Oxidase / Q000378:metabolism*; D006735:Horseradish Peroxidase / Q000378:metabolism*; D006801:Humans; D008297:Male; D008863:Microspheres; D010969:Plastics
D000588:Amines; D001786:Blood Glucose; D004800:Enzymes, Immobilized; D010969:Plastics; D005949:Glucose Oxidase; D006735:Horseradish Peroxidase