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Method for detecting immunoglobulin IgG based on monodisperse polymer and application thereof

An immunoglobulin and polymer technology, which is used in measurement devices, material separation, and analysis of materials. It can solve problems such as poor peak shape, inaccurate data, and inability to know the recovery rate of the method. less distracting effect

Active Publication Date: 2020-11-03
纳谱分析技术(苏州)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004]However, this method has the following defects: 1. The matrix of most foods is relatively complex. Since there is no pretreatment, the miscellaneous protein will peak together with IgG, 2. It is impossible to know the recovery rate of the method, because there may be a drop in detection sensitivity due to the incomplete elution of the filler after the IgG is adsorbed; 3. The peak shape of the detection is not good, due to different processes 4. The eluent cannot completely elute IgG, and the blank injection will also have IgG target substance peaks, resulting in inaccurate detected data

Method used

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  • Method for detecting immunoglobulin IgG based on monodisperse polymer and application thereof
  • Method for detecting immunoglobulin IgG based on monodisperse polymer and application thereof

Examples

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Embodiment 1

[0061] This embodiment provides a method for detecting immunoglobulin IgG based on a monodisperse polymer, the method comprising the following steps:

[0062] (1) Affinity Chromatography Solid Phase Extraction Purified Samples

[0063] Packing: Select monodisperse polyacrylate microspheres as matrix coupling ProteinG, and use it as filler, take 1mL filler and 3mL polypropylene tube to pack into solid-phase extraction column; Wherein, the monodisperse polyacrylate microsphere The particle size is 65μm;

[0064] Activation: Use 5mL double distilled water and 10mL 0.05mol / L PBS buffer solution with pH6.5 to activate the small column in sequence;

[0065] Sample loading: take 1mL sample and slowly add it to the solid phase extraction column;

[0066] Rinse: Use 5mL of 0.05mol / L PBS buffer solution with a pH of 6.5 as the eluent to rinse the solid-phase extraction column;

[0067] Elution: Use 3mL sodium acetate-acetic acid buffer solution (pH 3.0) for elution, and collect all e...

Embodiment 2

[0073] The difference from Example 1 is that the particle size of the monodisperse polymer microspheres is 90 μm, and other conditions are consistent with Example 1.

Embodiment 3

[0075] The difference from Example 1 is that the material of the monodisperse polymer microspheres is polystyrene, and the rest of the conditions are consistent with Example 1.

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Abstract

The invention provides a method for detecting immunoglobulin IgG based on a monodisperse polymer and application thereof. The method comprises the following steps: (1) transferring a sample to be detected into a solid-phase extraction column, performing leaching after affinity chromatography solid-phase extraction, and performing eluting to obtain an eluent, wherein filler of the solid-phase extraction column comprises monodisperse polymer microspheres and protein G connected to the monodisperse polymer microspheres through covalent bonds; and (2) analyzing the eluent by using high performanceliquid volume exclusion chromatography to obtain the content of immunoglobulin IgG in the sample to be detected. According to the method, monodisperse polymer microspheres are used as a binding matrix of protein G, IgG of various subtypes can be captured, other proteins in a sample are not captured, the sensitivity and specificity are good, quantitative analysis is carried out by adopting high performance liquid volume exclusion chromatography, and an obtained chromatogram is good in peak pattern, free of bifurcation and trailing and high in sensitivity and specificity. The method is suitablefor analysis of complex matrix samples such as food and health care products.

Description

technical field [0001] The invention relates to the field of sample analysis and measurement, in particular to a method for detecting immunoglobulin IgG based on a monodisperse polymer and its application, in particular to a method for detecting immunoglobulin IgG based on a monodisperse polymer affinity chromatography solid-phase extraction-liquid chromatography Methods and applications of globulin IgG. Background technique [0002] Immunoglobulin (Ig) is a group of proteins with antibody activity, including IgG, IgA, IgM, IgD and IgE. In recent years, more and more food and health care products have been added with IgG-based immunoglobulin, which has various functions such as regulating human metabolism, enhancing body immunity, and anti-infection. [0003] At present, IgG detection methods mainly include enzyme-linked immunosorbent assay, high performance liquid chromatography, immunocolloidal gold method and capillary electrophoresis, etc. Among them, high performance l...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06G01N30/34G01N30/86
CPCG01N30/02G01N30/06G01N30/34G01N30/8679G01N2030/062
Inventor 张勇侯玉
Owner 纳谱分析技术(苏州)有限公司
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