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A molecular weight detection method and application of synthetic antigen

A detection method and molecular weight technology, which is applied in the direction of measuring devices, material separation, and analysis of materials, can solve the problems of high detection cost, low specificity, and difficulty in popularization, and achieve simple operation, low difficulty in data processing, and reliable detection results Effect

Active Publication Date: 2019-04-16
广东省药品检验所
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

UV spectrophotometry estimates the coupling ratio by simple summation of UV absorption, and its specificity is not strong; SDS polypropylene gel electrophoresis can roughly estimate the purity and molecular weight of antigen synthesis through mark marking, but its accuracy Poor; liquid mass spectrometry can accurately determine the molecular weight and confirm the purity, but its detection cost is high and requires professional analysis of the measured spectrum to obtain accurate results. This method requires both equipment and technical personnel. High, often generally not easy to popularize
Currently available mass spectrometry methods for molecular weight detection include Q-TOF and MALDI-TOF. The inventor once used Q-TOF to detect the molecular weight of antigens. Data analysis shows that this method requires high purity of the substance to be tested, such as the BSA reference substance Accurate determination can be carried out, but the antigen samples prepared by this project team cannot be analyzed
The reason is that the antigen components prepared by this method are non-purified substances, and contain a series of similar components at the same time, and the typical "five-finger peaks" of the Q-TOF spectrum cannot be obtained, so this method cannot be applied in this project
[0005] In summary, in the research and application of synthetic antigens, there is a lack of a simple and accurate method for detecting the molecular weight of synthetic antigens. The method of the present invention will make up for the shortcomings of the prior art and provide technical reference for the research and application of antigens

Method used

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  • A molecular weight detection method and application of synthetic antigen
  • A molecular weight detection method and application of synthetic antigen
  • A molecular weight detection method and application of synthetic antigen

Examples

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Effect test

Embodiment 1

[0035] Method for Determining the Molecular Weight of Phenolphthalein Antigen

[0036] 1. Instruments and materials

[0037] 1.1 Instruments and equipment: Agilent 1100 series liquid chromatograph; Mollipore pure water system;

[0038] Column: Protein BEH SEC (7.8mm×150mm).

[0039] 1.2 Control substances: human serum IgG (≥95%, SDS-PAGE, SLBK8678V, SIGMA-ALDRICH); horse myoglobin (≥90%, SDS-PAGE, SLBK8678V, SIGMA-ALDRICH); serum albumin (bovine) ( BSA, batch number: 140619-201421, China National Institutes for Food and Drug Control).

[0040] 1.3 Reagents: Disodium hydrogen phosphate and phosphoric acid are analytically pure.

[0041] 2. Detection method

[0042] Chromatographic conditions and system suitability: the chromatographic column is Protein BEH SEC (7.8mm×150mm); mobile phase is 100mM phosphate buffer (adjust pH 6.8 with phosphoric acid); flow rate is 0.8mL / min; column temperature is 25°C; detection wavelength is 280nm.

[0043] Preparation of the refer...

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Abstract

The invention discloses a simple detection method of the molecular weight of a synthetic antigen. A standard curve is drafted through gel chromatography, and the standard curve is used to detect the molecular weight of antigen molecules. The detection method of the molecular weight of the synthetic antigen can be applied to detect the synthetic antigen, antibodies and protein macromolecular substances, and the detected molecular weight range is 16-150 kDa. Coupling proportions of micro-molecules and proteins in the synthesis process can be reckoned by detecting the molecular weight of the synthetic antigen and combining the molecular weights of the coupled proteins and micro-molecules. Compared with present detection methods, the method disclosed in the invention has the advantages of simplicity in operation, extremely small data processing difficulty, and reliable detection result. The method can be used to separate substance components with different molecular weights and calculate the molecular weights, and is especially suitable for detecting mixed macro-molecular substances.

Description

technical field [0001] The invention relates to a method for detecting the molecular weight of a synthetic antigen and determining small molecule compounds. Background technique [0002] Antigen is a kind of substance that can stimulate the immune system of the body to cause an immune response, produce antibodies and sensitized lymphocytes, etc., and can specifically bind and react with corresponding antibodies or sensitized lymphocytes in vivo or in vitro . Antigens in the body can stimulate the body to generate an immune response to generate specific antibodies. Since antibodies and antigens are specifically combined, ELISA or colloidal gold immunochromatography using this principle is one of the commonly used methods in clinical practice. [0003] For small molecule compounds with a molecular weight of less than 10,000, they are often not immunogenic, and it is difficult to cause an immune response in vivo. In order to obtain antibodies to such small molecular compounds...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/02
CPCG01N30/02
Inventor 杨立伟邱蕴绮欧国灯梁祈杨鹏博杨松林
Owner 广东省药品检验所
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