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Preparation method of glycated albumin calibrator

A technology for glycosylated albumin and calibrator, which is applied in the field of medical testing and can solve the problems of deviation between the measured value of GA and the true value, long preparation time, and fluctuation of GA concentration.

Active Publication Date: 2015-07-08
浙江夸烨生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The traditional GA calibrator is made of mixed human serum, which has the risk of being contaminated by HIV or HPV. The glucose in the serum will also react with albumin to generate GA, which will fluctuate the concentration of GA.
Li Jiaqi et al. (Tianjin Pharmacy, 1997,9(3):7-8.), Liu Yu et al. (Shanghai Medical Laboratory Journal, 1998,13(4):233-234.), Ningbo Meikang Biotechnology Co., Ltd. (patent Application number: CN201210572923.5) have reported the method of mixing glucose and albumin in water phase, glycosylation reaction for 7-30 days and dialysis purification for 2-4 days to prepare GA calibrator or quality control product, according to Although there is no free glucose in the glycosylated albumin solution prepared by this method, due to the long preparation time, part of the glycosylated albumin decomposes to produce glycosylated polypeptides, glycosylated amino acids and other substances. Remove glycosylated polypeptides, glycosylated amino acids and other substances, but during the long-term dialysis process, some glycosylated albumin decomposes to produce glycosylated polypeptides and glycosylated amino acids. These substances participate in the color reaction of FAOD colorimetry, making the measured value of GA and the true value There is a clear discrepancy between

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] (1), preparation containing 1,2-propanediol concentration is 50ml / L, the concentration of sodium chloride is 9g / L, the concentration of EDTA is 0.5mmol / L, the concentration of sodium azide is 0.2g / L, the concentration of glucose is 20ml of 20mmol / L pH 7.0 phosphate buffer solution of 20g / L, add 0.4g of recombinant human serum albumin to make the concentration 20g / L, and react at 25°C for 12 hours;

[0029] (2), the solution prepared by step (1) is transferred to the tangential flow filtration system, and the modified polyethersulfone membrane with a molecular weight cut-off of 60KD is selected, and the pump speed is 50ml / min; Stop the concentration operation when the concentration multiple is reached; carry out the elution operation, use 20mmol / L pH7.0 phosphoric acid containing sodium chloride concentration of 9g / L, EDTA concentration of 0.5mmol / L, and sodium azide concentration of 0.2g / L Salt buffer is used for elution, and the elution operation is stopped when the el...

Embodiment 2

[0034] (1), preparation containing 1,3-propanediol concentration is 100ml / L, the concentration of sodium chloride is 9g / L, the concentration of EDTA is 2.5mmol / L, the concentration of sodium azide is 1.0g / L, the concentration of glucose is 20ml of 35g / L 100mmol / L pH 7.5 phosphate buffer, add 0.7g of recombinant human serum albumin to make the concentration 35g / L, and react at 30°C for 24 hours;

[0035] (2), the solution prepared by step (1) is transferred to the tangential flow filtration system, and the modified polyethersulfone membrane with a molecular weight cut-off of 60KD is selected, and the pump speed is 65ml / min; Stop the concentration operation when the concentration is multiple; carry out the elution operation, use 100mmol / L pH 7.5 phosphate containing sodium chloride concentration of 9g / L, EDTA concentration of 2.5mmol / L, and sodium azide concentration of 1.0g / L The buffer solution is used for elution, and the elution operation is stopped when the elution multiple...

Embodiment 3

[0040] (1), the preparation contains 1,3-butanediol concentration of 200ml / L, the concentration of sodium chloride is 9g / L, the concentration of EDTA is 5mmol / L, the concentration of sodium azide is 2.0g / L, the concentration of glucose 20ml of 50g / L 200mmol / L pH 8.0 phosphate buffer, add 1.0g of recombinant human serum albumin to make the concentration 50g / L, and react at 37°C for 36 hours;

[0041] (2), the solution prepared by step (1) is transferred to the tangential flow filtration system, and the modified polyethersulfone membrane with a molecular weight cut-off of 60KD is selected, and the pump speed is 80ml / min; Stop the concentration operation when the concentration is multiple; carry out the elution operation, use 200mmol / L pH 8.0 phosphate buffer containing sodium chloride concentration of 9g / L, EDTA concentration of 5mmol / L, and sodium azide concentration of 2.0g / L solution for elution, and stop the elution operation when the elution multiple reaches 20 times;

[0...

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PUM

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Abstract

The invention discloses a preparation method of a glycated albumin calibrator. The preparation method includes following steps: (1) preparing a phosphate buffer liquid being 7.0-8.0 in pH, which comprises certain amounts of a glycosylation accelerator, sodium chloride, EDTA, sodium azide and glucose; (2) adding recombination human serum albumin and performing a glycosylation reaction for a certain time to prepare a solution; (3) transferring the solution to a tangential flow filtering system to remove glycosylated polypeptide, glycosylated amino acids and glucose; (4) diluting the filtered solution to a required concentration of the calibrator; and (5) adding an excipient, and packaging the calibrator separately and freeze-drying the calibrator. According to the method, the prepared glycated albumin calibrator is free of the glycosylated polypeptide, the glycosylated amino acids and the glucose. The method is suitable for mass production of the glycated albumin calibrator.

Description

technical field [0001] The invention belongs to the technical field of medical testing, and in particular relates to a preparation method of a glycated albumin calibrator. Background technique [0002] Glycated serum albumin (GA) is the product of serum albumin glycosylated by glucose, that is, the ε-amino group on the lysine residue of serum albumin is glycosylated. The half-life of glycosylated albumin is short, and the determination of the concentration of glycosylated albumin can effectively reflect the average blood sugar level of the patient in the past 2 to 3 weeks, and is not affected by temporary blood sugar concentration fluctuations. Glycosylated amino acid oxidase (FAOD) colorimetric method is currently the main method for the determination of GA. The traditional GA calibrator is made of mixed human serum, which has the risk of being contaminated by HIV or HPV, etc. The glucose coexisting in the serum will also continuously react with albumin to generate GA, cau...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/28
Inventor 连国军
Owner 浙江夸烨生物科技有限公司
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