Method for detecting immune globulin IgM 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: 2010-07-21
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 IgM in human milk, which solves the problem of low detection rate in the detection of human milk immunoglobulin IgM 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 30°C, using an uncoated quartz capillary column with a diameter of 60 μm and a length of 400 mm, an ultraviolet detector, pressure injection, a pressure of 0.5 psi, a time of 15 s, and a working voltage of 25 kV.

[0021] Establishment of 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 IgM standard substances. After shaking well, use high-pressure capillary electrophoresis to analyze. The peak areas after integration are 283561, 132651, 72354, 34215, and 738 respectively. The standard curve is obtained by using software. The linear regression correlation coefficient is 0.9986, and the minimum detection limit is 2ng. The content is 0.01mg / ml, the standard sample recovery rate reaches 91%, and the repeat rate can reach 96%. The above data are within the scope of the intended effect.

[0022] Processing sample ...

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 psi, a time of 1 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 IgM standard. After shaking well, use high-pressure capillary electrophoresis to analyze. The peak areas after integration are 285641, 152421, 76214, 38654, and 1734. The standard curve is obtained by using software. The linear regression correlation coefficient is 0.9986, and the minimum detection limit is 2ng. The output content is 0.02mg / ml, the standard sample recovery rate reaches 93%, and the repeat rate reaches 96%.

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

Embodiment 3

[0034]The capillary electrophoresis temperature was controlled at 35°C, using an uncoated quartz capillary column with a diameter of 65 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 27 kV.

[0035] Establishment of the 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 IgM standard substances. After shaking well, use high-pressure capillary electrophoresis analysis. The peak areas after integration are 554712, 262514, 69524, 14235, and 6354 respectively. The standard curve is obtained by using software. The linear regression correlation coefficient is 0.9994, 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, 7mo...

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Abstract

The invention relates to a method for detecting human milk ingredient, in particular to a method for detecting immune globulin IgM content in human milk, comprising: (1) dissolving a human immune globulin IgM standard substance into sample buffer solution to prepare to 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 solution to obtain a detection result, 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; (3) comparing the detection result of the sample with the standard curve to obtain the human immune globulin IgM 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 the content of immunoglobulin IgM in human milk, belonging to the technical field of detection. Background technique [0002] Human immunoglobulin IgM is another important protein in human serum, accounting for 5%-10% of the total serum immunoglobulin, the serum concentration is about 1mg / ml, the content in human milk is 20-120mg / L, and the molecular weight is 150~ 180KDa. IgM is the earliest antibody synthesized and secreted during individual development, and the fetus in the late embryonic development can produce IgM, which can improve the body's immunity and prevent infection. [0003] At present, the main methods for detecting human immunoglobulin IgM at home and abroad are ELISA, protein chip and SDS electrophoresis. These methods all have the disadvantages of complex operation, long time and low accuracy. In view of the ...

Claims

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Application Information

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IPC IPC(8): G01N27/447G01N27/453G01N1/28
Inventor 孙国庆康小红刘卫星
Owner INNER MONGOLIA MENGNIU DAIRY IND (GRP) CO LTD
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