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Enzyme-labeled secondary antibody compound and preparation method thereof

An enzyme-labeled secondary antibody and complex technology, applied in the field of immunoassay, can solve the problems of limited number of enzymes and antibodies that can be labeled, insufficient sensitivity, and low penetrating power, and can reduce the hydrodynamic volume, improve sensitivity, and improve penetration. The effect of penetration

Active Publication Date: 2020-08-18
河南赛诺特生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, this method is not perfect, and its main technical defects are: (1) the method adopts a dextran chain polymer backbone, which has a loose structure and a huge hydrodynamic volume, thereby causing great steric hindrance, making the polymer less permeable to the nuclear membrane
Therefore, the performance of this polymer for the detection of various types of antigens is uneven, especially in the detection of low-expressed antigens in some nuclei; (2) the method is to directly label enzymes and intact antibodies On the backbone, due to the limited surface area on the backbone, the number of enzymes and antibodies that can be labeled is limited, so the sensitivity cannot be further improved

Method used

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  • Enzyme-labeled secondary antibody compound and preparation method thereof
  • Enzyme-labeled secondary antibody compound and preparation method thereof
  • Enzyme-labeled secondary antibody compound and preparation method thereof

Examples

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Embodiment 1

[0065] The enzyme-labeled secondary antibody complex of this example includes hyperbranched aminated polydimethylhydroxypropionate Boltorn TM H40 and horseradish peroxidase, horseradish peroxidase linked to hyperbranched aminated polydimethylhydroxypropionate Boltorn via diethyl squarylate TM On H40, horseradish peroxidase is linked with goat anti-mouse secondary antibody IgG-Fab' fragment through diethyl squarylate.

[0066] Hyperbranched Aminated Polydimethylhydroxypropionate Boltorn TM There are amino groups that can be coupled with coupling agents at the end and middle of the branched chain; aminated Boltorn TM The structural diagram of the series of products is as follows: Figure 4 shown.

[0067] The schematic diagram of the immunoassay principle of the enzyme-labeled secondary antibody complex in this embodiment is as follows Figure 5 As shown, enzyme and antibody fragments are layered on the surface of the hyperbranched aminated polydimethylhydroxypropionate ba...

Embodiment 2

[0069] The enzyme-labeled secondary antibody complex of this example includes hyperbranched aminated polydimethylhydroxypropionate Boltorn TM H40 and horseradish peroxidase, horseradish peroxidase linked to hyperbranched aminated polydimethylhydroxypropionate Boltorn via diethyl squarylate TM On H40, horseradish peroxidase is linked with goat anti-rabbit secondary antibody IgG-Fab' fragment through diethyl squarylate.

Embodiment 3

[0071] The enzyme-labeled secondary antibody complex of this example includes hyperbranched aminated polydimethylhydroxypropionate Boltorn TM H40 and horseradish peroxidase, horseradish peroxidase linked to hyperbranched aminated polydimethylhydroxypropionate Boltorn via diethyl squarylate TM On H40, goat anti-mouse IgG-F(ab')2 fragment (Abcam, SKU: ab98634) is linked to horseradish peroxidase via diethyl squarylate.

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Abstract

The invention relates to an enzyme-labeled secondary antibody compound and a preparation method thereof, and belongs to the technical field of immunoassay. The enzyme-labeled secondary antibody compound comprises a polymer skeleton and a labeling enzyme. The labeling enzyme is connected to the polymer skeleton through a coupling agent, and a secondary antibody is connected to the labeling enzyme through a coupling agent; wherein the polymer skeleton is provided with a branched chain end part and a middle branch structure connected with the branched chain end part, and the labeling enzyme is connected to the branched chain end part and the middle branch structure of the polymer skeleton through a coupling agent; and the polymer skeleton is a hyper-branched polymer skeleton. The branched chain end part and the middle branch structure of the polymer skeleton can be connected with a coupling agent, so that the sensitivity of the enzyme-labeled secondary antibody compound is favorably improved, and the accumulation mode of enzyme and antibody is favorable for improving the polymerization degree of enzyme and antibody in the polymer, so that the sensitivity of the enzyme-labeled secondary antibody compound in immunoassay is greatly improved.

Description

technical field [0001] The invention relates to an enzyme-labeled secondary antibody complex and a preparation method thereof, belonging to the technical field of immune analysis. Background technique [0002] Immunoassay is a method for detecting protein biomarkers in samples through the principle of "antibody-antigen specific binding". Due to its high sensitivity and specificity, immunoassay can be widely used in the qualitative and quantitative analysis of various biological samples. Immunoassay methods include: ELISA, Western-Blot, Immunohistochemistry, Lateral Flow Assay, Protein Array, Cellular Immunoassay ( Immunocytochemistry) and so on. Among them, "enzyme-labeled secondary antibody complex" is widely used in the detection of biomarkers in the field of immunoanalysis, and its application fields include immunohistochemistry, enzyme-linked immunosorbent immunoassay, cell immunity and western blotting, etc. "enzyme-labeled secondary antibody complex" The principle i...

Claims

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Application Information

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IPC IPC(8): G01N33/535
CPCG01N33/535
Inventor 刘畅梁永波牛银银刘玲玲齐华
Owner 河南赛诺特生物技术有限公司
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