Method for detecting content of immunoglobulin IgG in human milk

An immunoglobulin and human detection technology, applied in the field of detection, can solve problems such as low detection rate, achieve high detection rate, prevent adsorption, and reduce costs

Inactive Publication Date: 2010-08-04
INNER MONGOLIA MENGNIU DAIRY IND (GRP) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] The present invention also provides a method for detecting IgG content in human milk, which...

Method used

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] The capillary electrophoresis temperature was controlled at 25°C, using an uncoated quartz capillary column with a diameter of 50 μm and a length of 500 mm, an ultraviolet detector, pressure injection, a pressure of 0.1 psi, a time of 15 s, and a working voltage of 15 kV.

[0021] Establishment of the standard curve: Human milk immunoglobulin IgG standard products were used to prepare standard solutions with concentrations of 4 mg / ml, 2 mg / ml, 1 mg / ml, 0.5 mg / ml, and 0.025 mg / ml, respectively. After shaking well, use high-pressure capillary electrophoresis analysis. The peak areas after integration are 65214, 35621, 15201, 7685, and 434 respectively. Using the software, the linear regression correlation coefficient of the standard curve is 0.9965, the minimum detection limit is 2ng, and the minimum detection content It is 0.02mg / ml, the recovery rate of the standard sample is 91%, and the repeatability is 95%.

[0022] Sample buffer configuration method: add 40mmol / L di...

Embodiment 2

[0027] The capillary electrophoresis temperature was controlled at 40°C, using an uncoated quartz capillary column with a diameter of 75 μm and a length of 600 mm, an ultraviolet detector, pressure injection, a pressure of 1.0 psi, a time of 1 s, and a working voltage of 25 kV.

[0028] Establishment of the standard curve: Human milk immunoglobulin IgG standard products were used to prepare standard solutions with concentrations of 4 mg / ml, 2 mg / ml, 1 mg / ml, 0.5 mg / ml, and 0.025 mg / ml, respectively. After shaking well, use high-pressure capillary electrophoresis to analyze. The peak areas after integration are 54721, 23154, 13265, 5124, and 211, respectively. The standard curve is obtained by using software. The linear regression correlation coefficient is 0.9942, and the minimum detection limit is 1ng. The output content is 0.01mg / ml, and the standard sample recovery rate reaches 92%.

[0029] Processing sample buffer configuration method: add 60mmol / L ethylenediaminetetraace...

Embodiment 3

[0034] The capillary electrophoresis temperature was controlled at 30°C, using an uncoated quartz capillary column with a diameter of 60 μm and a length of 500 mm, an ultraviolet detector, pressure injection, a pressure of 0.8 psi, a time of 6 s, and a working voltage of 25 kV.

[0035] Establishment of the standard curve: Human milk immunoglobulin IgG standard products were used to prepare standard solutions with concentrations of 8 mg / ml, 4 mg / ml, 1 mg / ml, 0.25 mg / ml, and 0.01 mg / ml, respectively. After shaking well, use high-pressure capillary electrophoresis to analyze. The peak areas after integration are 125412, 61245, 13658, 3956, and 1532 respectively. The standard curve is obtained by using software. The linear regression correlation coefficient is 0.9994, and the minimum detection limit is 1ng. The output content is 0.01mg / ml, and the standard sample recovery rate reaches 95%.

[0036] Processing sample buffer configuration method: add 50mmol / L ethylenediaminetetraacet...

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Abstract

The invention relates to a method for detecting human milk ingredients, in particular to a method for detecting immunoglobulin IgG content in human milk, comprising: (1) dissolving a human immunoglobulin IgG standard substance into sample buffer solution to prepare and obtain multiple standard solutions with different contents of the standard substance, using a high voltage capillary electrophoresis analyser to respectively detect and analyze the standard solutions to obtain detection results, and drawing a standard curve according to the concentration of the standard solution and the corresponding detection result; (2) after a human milk sample is centrifuged, utilizing the high voltage capillary electrophoresis analyser to detect the sample; and (3) comparing the detection result of the sample with the standard curve to obtain the human immunoglobulin IgG content in human milk sample. The invention has the characteristics of accurate detection result, high repeatability and low cost.

Description

technical field [0001] The invention relates to a method for analyzing and detecting protein in human milk, in particular to a method for detecting IgG content in human milk, belonging to the technical field of detection. Background technique [0002] Human immunoglobulin IgG is the main component of human serum immunoglobulin, accounting for 70-75% of the total immunoglobulin, the content in human milk is 10-340mg / L, the molecular weight is about 150KDa, and the sugar content is 2-3%. IgG molecules consist of 4 peptide chains. Among them, the peptide chain with a molecular weight of 25,000 (23kDa) is called the light chain (L chain), and the peptide chain with a molecular weight of 50,000 (50-60kDa) is called the heavy chain (H chain). The light chain and the heavy chain are linked by a disulfide bond (-S-S-). Human immunoglobulin IgG has antibacterial, antitoxic and antiviral effects. [0003] At present, the main methods for detecting human immunoglobulin IgG at home a...

Claims

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Application Information

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IPC IPC(8): G01N27/447
Inventor 孙国庆康小红刘卫星
Owner INNER MONGOLIA MENGNIU DAIRY IND (GRP) CO LTD
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