Test strip and method for detecting urine A beta amyloid protein

An amyloid and test strip technology, which is applied in biological tests, measuring devices, material inspection products, etc., can solve the problems of small target antigen, difficult to reach, high sensitivity and specificity, etc.

Active Publication Date: 2020-09-11
HUNAN QIANKANG TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The target antigen is too small, only 4.3kD, and is prone to non-specific binding with other proteins, so it is difficu...

Method used

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  • Test strip and method for detecting urine A beta amyloid protein
  • Test strip and method for detecting urine A beta amyloid protein
  • Test strip and method for detecting urine A beta amyloid protein

Examples

Experimental program
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Embodiment Construction

[0025] 1. Production and purification of anti-Aβ monoclonal antibody

[0026] A commercial mouse hybridoma cell line (BGM02) was purchased from Ab-Mart Pty Ltd to produce mouse anti-Aβ monoclonal antibody. Other commercially available monoclonal antibody clones include 1E8, 4G8, W0-2, 6E10, etc. can also be used alone or in combination. The hybridoma cells were thawed at 45°C using air-permeable hydrophobic cell culture flasks, and 1% glutamine, 1% streptomycin / penicillin, and 20% fetal bovine serum (FBS) were added to the hybridoma cell serum-free medium ( % refers to the mass percentage of the component in the serum-free medium of hybridoma cells).

[0027] When the culture medium turned yellow, the passage cells were carried out, and the ratio of serum in the passage cells decreased from 20%, 10%, 5%, 2% to the serum-free medium (0%), and the cells could survive. This process usually takes a month. Do not aspirate the supernatant until the serum has been consumed.

[00...

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PUM

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Abstract

The invention discloses a test strip and method for detecting urine A beta amyloid protein. The test strip for detecting the urine A beta amyloid protein comprises a PVC bottom plate (7), wherein a sample pad (1), a conjugated pad (2), a chromatography pad (3) and a water absorption pad (4) which are sequentially overlapped are paved on the PVC bottom plate (7); the conjugated pad (2) is coated with colloidal gold particles conjugated with a monoclonal antibody; a detection line (5) is arranged on one side, close to the conjugated pad (2), of the chromatography pad (3); a contrast control line(6) is arranged on one side, close to the water absorption pad (4), of the chromatography pad (3); the detection line (5) is coated with an A beta amyloid protein binding polymer; and the contrast control line (6) is coated with a goat anti-mouse IgG polyclonal antibody. The method is high in sensitivity, has a detection limit of 40 pg, is rapid and economical, is suitable for conventional clinical pathological examination, is suitable for general screening of a large number of people and self-screening of home terminal users, is especially used for clinically assisting early diagnosis and pre-judgment of mild cognitive impairment (MCI), and predicts and judges whether a patient has high risk of developing MCI and AD.

Description

technical field [0001] The invention relates to a test strip and a method for detecting urine Aβ amyloid protein. Background technique [0002] Alzheimer's disease (AD) is the most common form of dementia, affecting more than 26 million people worldwide. We and others have found that a large proportion (30%) of cognitively normal older adults exhibit high retention of PET ligands in the brain and low levels of Aβ in the cerebrospinal fluid (CSF) 1-42 , these individuals are currently diagnosed with Alzheimer's disease in the preclinical latent stage. Mild cognitive impairment (MCI) is considered a prodromal stage of AD, and 40-60% of patients meeting MCI criteria will eventually develop clinical AD, accounting for about 5-25% per year. The main etiology of sporadic AD is the accumulation of Aβ and the failure of Aβ clearance. At present, the "amyloid cascade hypothesis" is generally supported in this research field, that is, the metabolic imbalance of Aβ in the brain is t...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/58G01N33/577G01N33/558
CPCG01N33/6896G01N33/587G01N33/577G01N33/558G01N2333/4709G01N2800/2821
Inventor 戴正乾顾柏俊
Owner HUNAN QIANKANG TECH CO LTD
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