Method for quickly purifying glycated albumin coarse solution

A technology for glycated albumin and albumin, which is applied in the field of albumin purification and can solve problems such as deviation of GA measured values

Active Publication Date: 2015-06-24
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

Selecting a semi-permeable membrane with a suitable pore size to perform 2-4 days of dialysis purification on the crude solution of glycated albumin is a conventional purification method, although in theory, the method of dialysis for a long enough time can remove glycated polypeptides, glycated amino acids, and glucose. However, during the long-term dialysis process, some glycosylated albumin decomposes to produce glycosylated polypeptides and glycosylated amino acids, so that there are still a small amount of glycosylated polypeptides and glycosylated amino acids in the dialysis-purified glycosylated albumin solution. Color reaction, so there is a certain deviation between the FAOD colorimetric method and the GA measurement value of the reference method HPLC method

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] A method for rapidly purifying a crude glycated albumin solution, comprising the following steps:

[0023] 1) Put 50ml of the crude glycated albumin solution to be purified into a refrigerated centrifuge, select the centrifugation temperature as 2°C, and the centrifugation speed as 3000rpm, and take 49ml of the supernatant solution after centrifugation for 10 minutes;

[0024] 2) The 49ml supernatant solution prepared in step (1) is transferred to the sample container of the tangential flow filtration system, the ultrafiltration membrane is arranged inside the hollow fiber membrane filter, the ultrafiltration membrane is a modified polyethersulfone membrane, The molecular weight cut off is 60KD, and the membrane area is 20cm 2 , set the target pump speed to 20ml / min, and the shear force to 2000s -1 . Close the filtrate clamp of the filtration circuit, first adjust the pump speed to half of the target flow rate (10ml / min), and then turn on the peristaltic pump to fill ...

Embodiment 2

[0028] A method for rapidly purifying a crude glycated albumin solution, comprising the following steps:

[0029] 1) Put 500ml of the crude glycated albumin solution to be purified into a refrigerated centrifuge, select the centrifugation temperature as 5°C, and the centrifugation speed as 5500rpm, and take 490ml of the supernatant solution after centrifugation for 10 minutes;

[0030] 2) The 490ml supernatant solution prepared by step (1) is transferred to the sample container of the tangential flow filtration system, the ultrafiltration membrane is arranged inside the hollow fiber membrane filter, the ultrafiltration membrane is a modified polyethersulfone membrane, The molecular weight cut off is 60KD, and the membrane area is 115cm 2 , set the target pump speed to 35ml / min, and the shear force to 3000s -1. Close the filtrate clamp of the filtration circuit, first adjust the pump speed to half of the target flow rate (17.5ml / min), turn on the peristaltic pump, and fill th...

Embodiment 3

[0034] A method for rapidly purifying a crude glycated albumin solution, comprising the following steps:

[0035] 1) Put 1000ml of the crude glycated albumin solution to be purified into a refrigerated centrifuge, select the centrifugation temperature as 8°C, and the centrifugation speed as 8000rpm, and take 980ml of the supernatant solution after centrifugation for 10 minutes;

[0036] 2) The 980ml supernatant solution prepared by step (1) is transferred to the sample container of the tangential flow filtration system, the ultrafiltration membrane is arranged inside the hollow fiber membrane filter, the ultrafiltration membrane is a modified polyethersulfone membrane, The molecular weight cut off is 60KD, and the membrane area is 790cm 2 , set the target pump speed to 50ml / min, and the shear force to 4000s -1 . Close the filtrate clamp of the filtration circuit, first adjust the pump speed to half of the target flow rate (25ml / min), and then turn on the peristaltic pump to ...

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Abstract

The invention discloses a method for quickly purifying a glycated albumin coarse solution. The method comprises the following steps: (1) performing low-temperature centrifugation on the glycated albumin coarse solution, and taking a supernatant solution; (2) transferring the supernatant solution to a tangent flow filtering system for concentration treatment to obtain a coarse concentrated glycated albumin coarse solution; (3) performing dilution treatment in the tangent flow filtering system to remove glycosylated peptides, glycated amino acid and glucose from the coarse concentrated glycated albumin coarse solution; and (4) diluting the purified glycated albumin coarse solution to the required concentration by a buffering solution. The glycated albumin coarse solution is purified by the method, so that small-molecular impurities such as glycosylated peptides can be quickly removed; the method has the characteristics of simplicity, convenience and quickness in operation and the like and is suitable for large-scale purification of the glycated albumin coarse solution.

Description

technical field [0001] The invention belongs to the technical field of albumin purification, and in particular relates to a method for quickly purifying a crude glycosylated albumin solution. Background technique [0002] Glycated albumin (GA) is formed by the non-enzymatic glycosylation reaction between glucose in the blood and the N-terminus of the albumin molecule. The measurement of glycosylated serum albumin can effectively reflect the average blood glucose level of the patient in the past 2 to 3 weeks, and is not affected by the time The effect of blood sugar concentration is a good indicator of blood sugar control in diabetic patients. The glycosylated amino acid oxidase (FAOD) colorimetric method is currently the main method for determining GA. In this method, a glycosylated albumin solution that does not contain glycosylated polypeptides, glycosylated amino acids, and glucose is required as a standard solution and a quality control solution for control testing and q...

Claims

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Application Information

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IPC IPC(8): G01N1/34
Inventor 连国军陶国锋
Owner 浙江夸烨生物科技有限公司
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