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Coupling method of fluorescent latex microspheres and protein

A fluorescent latex microsphere and coupling technology, applied in the field of biological analysis, can solve the problems of low coupling efficiency and coupling strength, low fluorescence value of the detection card, etc., to improve stability, avoid agglutination, and improve coupling efficiency and the effect of coupling strength

Pending Publication Date: 2020-04-28
BEIJING PEPMAGIC BIOTECH CORPORATION LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] For this reason, the embodiment of the present invention provides a coupling method of fluorescent latex microspheres and proteins to solve the problem of low fluorescence value of the detection card caused by the low coupling efficiency and coupling strength of fluorescent microspheres and proteins in the prior art

Method used

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  • Coupling method of fluorescent latex microspheres and protein
  • Coupling method of fluorescent latex microspheres and protein

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Embodiment 1

[0029] The present embodiment is a coupling method of fluorescent latex microspheres and cardiac troponin I monoclonal antibody, and the coupling method comprises the following steps:

[0030] (a) Dilute the fluorescent latex microspheres with MES buffer to a solid content of 0.8‰; then add an ethanol solution containing NHS and an ethanol solution containing EDC to the diluted fluorescent latex microsphere solution, mix well, and react at room temperature 25min, sonication, centrifugation at 10000r / min for 10min, collecting and collecting the activated fluorescent latex microspheres, adding MES buffer, ultrasonic treatment to obtain a fluorescent latex microsphere solution with a solid content of 0.08% of the fluorescent latex microspheres, wherein, The volume ratio of the volume ratio of the ethanol solution containing NHS and the ethanol solution containing EDC to the fluorescent latex microsphere solution is 1:26; the concentration of NHS in the ethanol solution containing ...

Embodiment 2

[0035] The present embodiment is a coupling method of fluorescent latex microspheres and cardiac troponin I monoclonal antibody, and the coupling method comprises the following steps:

[0036] (a) Dilute the fluorescent latex microspheres with MES buffer to a solid content of 0.8‰; then add an ethanol solution containing NHS and an ethanol solution containing EDC to the diluted fluorescent latex microsphere solution, mix well, and react at room temperature 25min, sonication, centrifugation at 10000r / min for 10min, collecting and collecting the activated fluorescent latex microspheres, adding MES buffer, ultrasonic treatment to obtain a fluorescent latex microsphere solution with a solid content of 0.08% of the fluorescent latex microspheres, wherein, The volume ratio of the volume ratio of the ethanol solution containing NHS and the ethanol solution containing EDC to the fluorescent latex microsphere solution is 1:28; the concentration of NHS in the ethanol solution containing ...

Embodiment 3

[0041] The present embodiment is a coupling method of fluorescent latex microspheres and cardiac troponin I monoclonal antibody, and the coupling method comprises the following steps:

[0042](a) Dilute the fluorescent latex microspheres with MES buffer to a solid content of 1.2‰; then add an ethanol solution containing NHS and an ethanol solution containing EDC to the diluted fluorescent latex microsphere solution, mix well, and react at room temperature 25min, sonication, centrifugation at 10000r / min for 10min, collecting and collecting the activated fluorescent latex microspheres, adding MES buffer, ultrasonic treatment to obtain a fluorescent latex microsphere solution with a solid content of 0.08% of the fluorescent latex microspheres, wherein, The volume ratio of the volume ratio of the ethanol solution containing NHS and the ethanol solution containing EDC to the fluorescent latex microsphere solution is 1:22; the concentration of NHS in the ethanol solution containing N...

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Abstract

The embodiment of the invention discloses a coupling method of fluorescent latex microspheres and protein. The method comprises the following steps: carrying out activation treatment and resuspensionon the fluorescent latex microspheres to obtain a fluorescent latex microsphere solution; diluting the protein by adopting a glycine solution, then adding the diluted protein into the fluorescent latex microsphere solution, carrying out ultrasonic reaction, then carrying out rotary uniform mixing reaction, and carrying out co-labeling on glycine and the protein; after the reaction is finished, adding confining liquid into the reaction liquid for confining, centrifuging, and collecting precipitates; and putting the precipitates into a glycine solution, and carrying out ultrasonic dispersion toobtain a fluorescent latex microsphere and protein coupled compound. According to the coupling method, coupling efficiency and coupling strength of the fluorescent latex microspheres and the protein can be improved so that surfaces of the microspheres are fully combined with binding sites of proteins, stability of the prepared compound can be effectively improved, and an agglutination phenomenon is avoided; in addition, sensitivity of fluorescence detection can be improved.

Description

technical field [0001] The embodiment of the present invention relates to the technical field of biological analysis, in particular to a coupling method of fluorescent latex microspheres and proteins. Background technique [0002] When detecting certain antigens or specific proteins, immunolabeling techniques are usually applied. Immunolabeling technology is a technology that marks the substance that is easy to measure and has high sensitivity to specific antigen or antibody protein, and shows the nature and content of the antigen or antibody in the reaction system through the enhanced amplification of these markers. [0003] Currently commonly used labels include colloidal gold, latex, fluorescein, fluorescent microspheres, enzymes, and radionuclides. However, in the existing process of labeling antibodies with fluorescent microspheres, due to unreasonable labeling methods, low coupling efficiency and coupling strength between fluorescent microspheres and antibodies leads ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/533G01N21/64
CPCG01N33/577G01N33/533G01N21/6428G01N21/6486
Inventor 管静波赵树民孙如石松传郭波
Owner BEIJING PEPMAGIC BIOTECH CORPORATION LTD
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