A method for detecting glucose using mercaptopropyl agarose spheres loaded with glucose oxidase and catalase
A technology of mercaptopropyl agarose spheres and glucose oxidase, applied in the field of glucose detection, can solve the problems of no diagnostic significance, high reaction conditions, and low dosage for diabetic patients, and achieve easy detection operation and observation, mild reaction conditions, The effect of color change is obvious
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Embodiment 1
[0062] Test strips (such as figure 1 ) preparation process:
[0063] a) First add 2ml of double distilled water to 0.2ml of dry mercaptopropyl agarose spheres Thiopropyl After soaking overnight in 6B (average size 90 μm), the mercaptopropyl agarose spheres swelled by water absorption, and their volume was about 3 times that of the dry state. The supernatant was discarded by centrifugation, and the tris(2-carboxyethyl)phosphine (TCEP) solution was reacted with the above-mentioned mercaptopropyl agarose spheres, glucose oxidase solution and catalase solution for 1 hour at room temperature, respectively, to make them double. Sulfur bonds open. The concentration of TCEP solution is 0.5mM, the solvent is 10mM Tris-HCl solution, the concentration of glucose oxidase solution and catalase solution is 50μg / ml, and the solvent is 10mM PBS (pH7.4); the enzyme activity of glucose oxidase is 100-250units / mg, the enzyme activity of catalase is ≥250units / mg.
[0064] b) The TCEP-treate...
Embodiment 2
[0069] Test strips (such as figure 1 ) preparation process:
[0070] a) First add 2ml of double distilled water to 0.2ml of dry mercaptopropyl agarose spheres Thiopropyl After soaking in 6B overnight, the mercaptopropyl agarose sphere swelled with water, and its volume was about 3 times that of the dry state. The supernatant was discarded by centrifugation, and the dithiothreitol (DTT) solution was reacted with the above-mentioned mercaptopropyl agarose spheres, glucose oxidase solution and catalase solution for 1 hour at room temperature to open the disulfide bonds. The concentration of DTT solution was 10 mM, the solvent was 10 mM Tris-HCl solution, the concentration of glucose oxidase solution and catalase solution was 30 μg / ml, and the solvent was 10 mM PBS (pH 7.4).
[0071] b) DTT-treated activated mercaptopropyl agarose spheres were washed 5 times with 10 mM PBS (pH 7.4) buffer, and the supernatant was removed. Then, 1.5 ml of DTT-treated activated glucose oxidase ...
Embodiment 3
[0076] Preparation process of enzyme-loaded mercaptopropyl agarose spheres:
[0077] a) First add 2ml of double distilled water to 0.2ml of dry mercaptopropyl agarose spheres Thiopropyl After soaking in 6B overnight, the mercaptopropyl agarose sphere swelled with water, and its volume was about 3 times that of the dry state. The supernatant was discarded by centrifugation, and the tris(2-carboxyethyl)phosphine (TCEP) solution was reacted with the above-mentioned mercaptopropyl agarose spheres, glucose oxidase solution and catalase solution for 1 hour at room temperature, respectively, to make them double. Sulfur bonds open. The concentration of TCEP solution was 0.5 mM, the solvent was 10 mM Tris-HCl solution, the concentration of glucose oxidase solution and catalase solution was 50 μg / ml, and the solvent was 10 mM PBS (pH 7.4).
[0078] b) The TCEP-treated activated mercaptopropyl agarose spheres were washed 5 times with 10 mM PBS (pH 7.4) buffer, and the supernatant was ...
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