Biosensor for synchronously detecting amyloid polypeptide monomers and aggregate and construction method and application thereof

An amyloid polypeptide and biosensor technology, applied in biological tests, instruments, measuring devices, etc., can solve problems such as high minimum concentration, and achieve the effects of low detection limit, improved sensitivity and strong specificity

Active Publication Date: 2016-06-29
CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In previous studies, although the amount of its polypeptide monomers and aggregates can be detected, the minimum concentration of its detection is relatively large, so the sensitivity of its detection needs to be improved

Method used

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  • Biosensor for synchronously detecting amyloid polypeptide monomers and aggregate and construction method and application thereof
  • Biosensor for synchronously detecting amyloid polypeptide monomers and aggregate and construction method and application thereof
  • Biosensor for synchronously detecting amyloid polypeptide monomers and aggregate and construction method and application thereof

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Embodiment 1

[0032] 1. The method for the biosensor constructed on the electrode, the steps are as follows:

[0033] At room temperature, modify 10 μM cDNA on the electrode for 16h, then wash with secondary water to remove the residual cDNA that is not modified, then place the electrode in MCH, the purpose of adding MCH is to seal the unreacted site on the electrode And make the modified cDNA in an upright structure, after being blocked in MCH for 1 hour, rinse with alcohol first, and then rinse with secondary water; then modify 10 μM DNA1 on the electrode, place it at room temperature for 2 hours, and then rinse with secondary water to remove Unreacted DNA1; then modify 10 μM DNA2-MB on the electrode, because methylene blue is easy to decompose when exposed to light, so from this step, all operations are carried out under dark conditions. After modification for 2 hours, use secondary water Rinse; then modify 10 μM DNA3-biotin on the electrode for 2h, and also wash with secondary water; th...

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Abstract

The invention discloses a biosensor for synchronously detecting amyloid polypeptide monomers and aggregate and a construction method and application thereof. The biosensor comprises an amyloid polypeptide aggregate aptamer containing a methylene blue probe, and antibody modified DNA. The construction method of the biosensor includes the steps that sulfydryl modified cDNA is fixed to an electrode through chemical bonding, after short chain DNA, the amyloid polypeptide aggregate aptamer containing the methylene blue probe and DNA connected with biotins are sequentially modified on the cNDA in a complementary mode, the object further reacts with streptavidin and an antibody connected with biotins, and then the biosensor is obtained. The construction method of the biosensor is simple, and the biosensor can be used for respectively or simultaneously detecting amyloid polypeptide monomers and aggregate, and has the advantages of being simpler in detection, low in detection limit, high in sensitivity and high in specificity.

Description

technical field [0001] The invention relates to a biosensor for synchronously detecting amyloid polypeptide monomers and aggregates, its construction method, and its application in detecting amyloid polypeptide monomers and aggregates, belonging to the field of electrochemical biosensors. Background technique [0002] Alzheimer's disease (AD) is a neurodegenerative disease with insidious onset and progressive development. Currently, according to the World Alzheimer's Disease Report, more than 35 million people worldwide are suffering from AD, and it is estimated that the number of people suffering from AD will double by 2030. Therefore, being able to prepare markers and detect AD becomes an urgent problem. Constructing a new type of electrochemical biosensor on the electrode surface can accurately measure the amount of marker Aβ1-40 monomer and aggregate. Although the amount of its polypeptide monomers and aggregates can be detected in previous studies, the minimum concent...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/327G01N33/68
CPCG01N27/3275G01N33/68
Inventor 邓春艳刘春燕
Owner CENT SOUTH UNIV
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