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Method for detecting immunoglobulin IgA content in human milk

A technology for immunoglobulin and human detection, applied in the field of detection, can solve the problems of low detection rate, achieve high detection rate, reduce cost, and prevent adsorption

Inactive Publication Date: 2010-08-18
INNER MONGOLIA MENGNIU DAIRY IND (GRP) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The invention also provides a method for detecting the content of immunoglobulin IgA in human milk, which solves the problem of low detection rate in the detection of human milk immunoglobulin IgA content in the prior art

Method used

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Examples

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 40 μm and a length of 400 mm, an ultraviolet detector, pressure injection, a pressure of 0.2 psi, a time of 20 s, and a working voltage of 20 kV.

[0021] Establishment of the standard curve: Standard solutions with concentrations of 4mg / ml, 2mg / ml, 1mg / ml, 0.5mg / ml, and 0.01mg / ml were prepared using human milk immunoglobulin IgA standard substances. After shaking well, use high-pressure capillary electrophoresis to analyze. The peak areas after integration are 25124, 12254, 6388, 3452, and 157 respectively. Use software to obtain a standard curve. The linear regression correlation coefficient is 0.9982, the minimum detection limit is 1ng, and the minimum detection content It is 0.01mg / ml, the recovery rate of the standard sample can reach 94%, and the repetition rate can reach 96%. The above data are within the scope of the intended effect.

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Embodiment 2

[0027] The capillary electrophoresis temperature was controlled at 45°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 1.2 psi, a time of 2 s, and a working voltage of 30 kV.

[0028] Establishment of standard curve: Standard solutions with concentrations of 4mg / ml, 2mg / ml, 1mg / ml, 0.5mg / ml and 0.025mg / ml were prepared using human milk immunoglobulin IgA standard. After shaking well, use high-pressure capillary electrophoresis analysis. The peak areas after integration are 22101, 10321, 5682, 2654, and 1542, respectively. The standard curve is obtained by using software. The linear regression correlation coefficient is 0.9982, and the minimum detection limit is 2ng. The output content is 0.02mg / ml, and the standard sample recovery rate reaches 94%.

[0029] Processing sample buffer configuration method: add 60mmol / L ethylenediaminetetraacetic acid, 8mol / L urea to 160mmol / L t...

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 standard curve: Standard solutions with concentrations of 8 mg / ml, 4 mg / ml, 1 mg / ml, 0.25 mg / ml, and 0.01 mg / ml were prepared using human milk immunoglobulin IgA standard. After shaking well, use high-pressure capillary electrophoresis analysis. The peak areas after integration are 51241, 24125, 6214, 1421, and 624 respectively. The standard curve is obtained by using software. The linear regression correlation coefficient is 0.9989, and the minimum detection limit is 2ng. The output content is 0.02mg / ml, and the standard sample recovery rate reaches 91%.

[0036] Processing sample buffer configuration method: add 50mmol / L ethylenediaminetetraacetic acid, 7mol / L urea to 150mmol...

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Abstract

The invention relates to a method for detecting the immunoglobulin IgA content in human milk, in particular to a detection method for detecting immunoglobulin IgA content in human milk. The method comprises the following steps: (1) dissolving human immunoglobulin IgA standards in sample buffer solutions, preparing multiple standard solutions with different standard contents, using a high-pressure capillary electrophoresis apparatus to respectively detect and analyze the standard solutions to obtain detection results, and drawing standard curves based on the concentrations of the standard solutions and the corresponding detection results; (2) centrifuging a human milk sample, and then detecting the sample by using the high-pressure capillary electrophoresis apparatus; and (3) comparing the detection result of the sample with the standard curves to obtain the human immunoglobulin IgA content in the 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 the content of immunoglobulin IgA in human milk, belonging to the technical field of detection. Background technique [0002] Human immunoglobulin IgA is one of the important components of human serum immunoglobulin, and its content in normal human serum is second only to IgG, accounting for 10-20% of serum immunoglobulin content. The relative molecular weight is 160KDa or 320KDa. IgA is divided into serotype and secretory type according to its immune function. Serum type IgA exists in the serum, its content accounts for about 85% of the total IgA, and has some functions of IgG and IgA. Secretory IgA exists in secretions, such as saliva, tears, colostrum, nasal and bronchial secretions, gastrointestinal fluids, urine, sweat, etc. IgA cannot pass through the placenta. There is no IgA antibody in neonatal serum, but secretory Ig...

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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