Quantitative determination method for saccharified protein

A technology for the quantitative determination of glycated protein, which is applied in the field of medical testing, can solve problems such as prolonged measurement time, increased background, and increased measurement error, and achieves the effects of convenient sampling, less pain for patients, and an objective percentage of glycated protein

Inactive Publication Date: 2009-07-29
SHANGHAI YESBIO BIO TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disadvantage of this method is that the background will definitely increase during the simultaneous incubation of the specific antibody enzyme conjugate and the uncoated solid phase
The result is often: the measurement time is prolonged and the measurement error is increased

Method used

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  • Quantitative determination method for saccharified protein
  • Quantitative determination method for saccharified protein
  • Quantitative determination method for saccharified protein

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0083] Example 1, the determination of glycated hemoglobin.

[0084] · Preparation of solid-phase antibodies:

[0085] Antibody A, mouse anti-human hemoglobin monoclonal antibody, American Abcam Company, model: AB401 Antibody A needs to be coated on a microwell plate (Sigma-Aldrich, M0156), and the antibody concentration is between 1.0 μg / ml and 10.0 μg / ml Between, coating buffer, 0.1M sodium bicarbonate buffer, pH 9.6. Coating volume: 200 microliters / microwell, overnight at room temperature. Then, 300 μl / well of blocking buffer (Sigma-Aldrich, C9483) was used to block overnight at room temperature. After drying, dry and store for later use.

[0086] · Preparation of antibody-labeled enzyme conjugate

[0087] Antibody B, mouse anti-human glycosylated hemoglobin monoclonal antibody, American Abcam Company, model: AB47149 Antibody C, mouse anti-human hemoglobin monoclonal antibody, American Abcam Company, model: AB402

[0088] Antibody B and Antibody C were each conjugated w...

Embodiment 2

[0132] Example 2, Determination of glycated albumin.

[0133] · Preparation of solid-phase antibodies:

[0134] Antibody A, goat anti-human serum albumin polyclonal antibody, American Abcam Company, model: AB19180, antibody A was coated on a microwell plate (Sigma-Aldrich, MO156), and the preparation method was the same as in Example 1.

[0135] · Preparation of antibody-labeled enzyme conjugate

[0136] Antibody B, mouse anti-human serum glycosylated albumin monoclonal antibody, American USBiological Company, model: A1327-55; Antibody C: mouse anti-human serum albumin monoclonal antibody, American Abcam Company, model: AB7793

[0137] Antibody B and Antibody C need to be conjugated with horseradish peroxidase (Sigma-Aldrich, P8415), respectively. Become an antibody-labeled enzyme conjugate. Conjugation method The preparation method is the same as in Example 1.

[0138] · Preparation of human serum albumin and human serum glycated albumin standards:

[0139] Human serum a...

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PUM

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Abstract

The invention provides a method for determining saccharified specific protein, using two groups of antibody pairs; one group is specific to saccharified protein and the other group is specific to total protein; the saccharified specific protein and the non-saccharified specific protein can be simultaneously determined, thus obtaining the ratio thereof by calculation. The high degree of specificity and affinity of the antibody group ensures the accuracy of tests, and shortens the test time.

Description

technical field [0001] The invention relates to the field of medical detection, in particular to a method for quantitatively measuring glycosylated proteins. Background technique [0002] 1. Clinical significance: [0003] Diabetes mellitus is a group of endocrine and metabolic diseases whose etiology and pathogenesis have not yet been fully understood. The current incidence rate is second only to cardiovascular and cerebrovascular diseases and tumors. In my country, the incidence rate of diabetes is 2-3%, and it is increasing at a rate of one thousandth. At present, the work of detecting blood glucose in patients has been widely carried out clinically. However, blood glucose measurement only represents the immediate blood glucose level, indicating the patient's physical condition at that time, and cannot be used as an indicator to evaluate the degree of disease control. [0004] Diabetes often causes retinal, renal, neurological and other lesions and various complication...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/543G01N33/72
Inventor 尤少方
Owner SHANGHAI YESBIO BIO TECH CO LTD
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