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Preparation method of glycosylated hemoglobin calibration quality control product

A technology of glycosylated hemoglobin and hemoglobin, which is applied in the field of medical testing, can solve the problems of biological safety of blood products, unfavorable industrialization, and rare samples, and achieve stable and controllable methods, which are conducive to industrial production and easy to obtain.

Pending Publication Date: 2022-01-21
上海瀚诺威生物科技有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, most of the HbA1c calibration quality control products use human whole blood as the raw material, but the acquisition of different concentration values ​​is random and uncontrollable, which is not conducive to industrialization
[0005] At present, human whole blood is mostly used to prepare glycated hemoglobin calibration quality control products. Blood products have biological safety problems, and blood samples of different concentrations are obtained randomly, which is not conducive to industrialization, and low-value and high-value samples are rare and difficult to obtain.

Method used

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  • Preparation method of glycosylated hemoglobin calibration quality control product
  • Preparation method of glycosylated hemoglobin calibration quality control product
  • Preparation method of glycosylated hemoglobin calibration quality control product

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Embodiment 1 prepares hemoglobin calibration standard

[0037] S1. Pretreatment of pig blood samples: collect fresh pig blood samples, mix them with the cleaning solution at a ratio of 1:1, mix well, centrifuge, remove the supernatant, and repeat three times to obtain pig red blood cells. The components of the cleaning solution are as follows:

[0038] Sodium chloride NaCl: 9g / L

[0039] CAA: 1%

[0040] S2, preparation of porcine hemoglobin: freeze-thaw the red blood cells obtained in S1 once at -20°C, add 1-10 times the volume of purified water, stir slowly at 2-8°C for 4-8 hours, and centrifuge to take the supernatant , to obtain porcine hemoglobin;

[0041] S3, glycosylation reaction: add the hemoglobin obtained in S2 to the glycosylation solution at a ratio of 1:10, and place it at 37°C for 3 days, and the composition of the glycosylation solution is as follows:

[0042] HEPES: 20mM

[0043] Sodium nitrite: 1g / L

[0044] CAA: 1%

[0045] Glucose: 50mM

[004...

Embodiment 2

[0061] Embodiment 2 prepares hemoglobin calibration standard

[0062] The difference from Example 1 is that the mass concentration of the preservative CAA is 0.1%, 2 mM ethylenediaminetetraacetic acid (EDTA) is added to the glycosylation fluid and the dialysate, and the rest of the conditions and test methods are the same as in Example 1.

[0063] S1. Pretreatment of pig blood samples: collect fresh pig blood samples, mix them with the cleaning solution at a ratio of 1:1, mix well, centrifuge, remove the supernatant, and repeat three times to obtain pig red blood cells. The components of the cleaning solution are as follows:

[0064] NaCl: 9g / L

[0065] CAA: 0.1%

[0066] S2, preparation of porcine hemoglobin: freeze-thaw the red blood cells obtained in S1 once at -20°C, add 1-10 times the volume of purified water, stir slowly at 2-8°C for 4-8 hours, and centrifuge to take the supernatant , to obtain porcine hemoglobin;

[0067] S3, glycosylation reaction: add the hemoglobi...

Embodiment 3

[0088] The optimization of embodiment 3 cleaning liquid addition proportions

[0089] The difference from Example 1 is that the ratio of fresh pig blood sample to cleaning solution is 1:2, and other conditions and testing methods are the same as in Example 1.

[0090] S1. Pretreatment of pig blood samples: collect fresh pig blood samples, mix them with the cleaning solution at a ratio of 1:2, centrifuge after mixing, remove the supernatant, and repeat three times to obtain porcine red blood cells. The components of the cleaning solution are as follows:

[0091] NaCl: 9g / L

[0092] CAA: 1%

[0093] S2, preparation of porcine hemoglobin: freeze-thaw the red blood cells obtained in S1 once at -20°C, add 1-10 times the volume of purified water, stir slowly at 2-8°C for 4-8 hours, and centrifuge to take the supernatant , to obtain porcine hemoglobin;

[0094] S3, glycosylation reaction: add the hemoglobin obtained in S2 to the glycosylation solution at a ratio of 1:10, and place...

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Abstract

The invention discloses a preparation method of a glycosylated hemoglobin calibration quality control product, and belongs to the technical field of medical examination. Fresh pig blood is used as a material for preparing the hemoglobin calibration quality control product, a high-value quality control product is prepared through cleaning, glycosylation reaction and dialysis purification, and calibration products with the percentage content being 5% and 12% and quality control products with the percentage content being 6% and 10% are obtained through prepared treating fluid. And the calibration quality control product covering the normal test range and the abnormal test range can be directly obtained at one time. Compared with the randomness of preparing the glycosylated hemoglobin calibration quality control product from human whole blood, the method is stable and controllable, the fresh pig blood is wide in source, low in cost and easy to obtain, the method is suitable for mass production, industrial production is facilitated, the yield of the hemoglobin calibration quality control product is effectively improved, and the biosafety problem does not exist. The hemoglobin calibration quality control product prepared by the method disclosed by the invention is good in stability, and has no obvious change before and after being stored for 12 months at 2-8 DEG C.

Description

technical field [0001] The invention relates to a preparation method of a glycosylated hemoglobin calibration quality control product, belonging to the technical field of medical testing. Background technique [0002] Glycated hemoglobin (Glycated hemoglobin GHb) is the product of the combination of hemoglobin in red blood cells and carbohydrates in serum, including HBA1a, HbA1b, and HbA1c, of which HbA1c accounts for about 70%. GHb is formed through a slow, continuous and irreversible glycation reaction, and its content depends on the blood sugar concentration and the contact time between blood sugar and hemoglobin, and has nothing to do with blood drawing time, whether the patient is fasting, whether insulin is used or not. [0003] At present, the clinical methods for determining the content of glycosylated hemoglobin can be divided into two categories according to different physical and chemical properties: one is based on the difference in charge between glycosylation a...

Claims

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Application Information

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IPC IPC(8): G01N1/28G01N1/38
CPCG01N1/28G01N1/38
Inventor 王晨谌林徐鑫帅
Owner 上海瀚诺威生物科技有限公司
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