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Method for determination of A-beta antibody and reagent kit therefor

A kit and antibody technology, applied in the field of medicine and health, can solve problems such as cumbersome methods, high requirements for equipment, and only antibodies can be compared, and achieve high accuracy, less interference, and simple methods

Inactive Publication Date: 2007-03-14
刘俊华
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, all these detection methods lack a unified standard product, can only compare antibody titers, cannot do quantitative detection, and require high equipment and cumbersome methods, which are not suitable for general clinical use

Method used

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  • Method for determination of A-beta antibody and reagent kit therefor

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] According to the following steps, we tested the Aβ in the serum of 10 cases of healthy persons (N1-10) and 4 cases of AD patients (A1-4). 1-42 Antibody levels were tested, and the concentration of phosphate buffered saline (PBS) in this example was 0.005 mol / L, and the pH value was 7.0.

[0054] 1) Coating: 96-well strips are made of polystyrene plates with good adsorption properties, and the difference between wells, plates and batches is CV1-42 Protein fragment 1.0ng / ul solution 50ul, 4 ℃ overnight, then discard the coating solution, the coated plate was washed twice with PBS containing 0.1% TW-20 by volume, pat dry, add 1% by weight Bovine serum albumin (BSA) and PBS with 1% by weight skimmed milk powder were blocked at room temperature for 2 hours, the blocking solution was discarded, and the coated plate was air-dried;

[0055] 2) Adding samples: Add the serum to be tested from the human body to each well of the coated strip, the amount of serum added is 50ul, and ...

Embodiment 2

[0065] According to the following steps, we tested the Aβ in the serum of the above 10 cases of healthy persons and 4 cases of AD patients 1-42 Antibody levels were tested repeatedly, and the concentration of the phosphate buffer in this example was 0.1 mol / L, and the pH value was 7.4.

[0066] 1) Coating: 48-well strips are made of polystyrene plates with good adsorption, and the difference between wells, plates and batches is less than 10%, and 0.05mol / L, PH9.6 carbonate buffer is used as the coating solution, add Aβ to each well 1-42 Protein fragment 100ng / ul solution 250ul, room temperature coating overnight, then discard the coating solution, wash the coated plate twice with PBS containing 0.1% TW-20 by volume, pat dry, add bovine serum containing 1% by weight Albumin (BSA) and PBS with 1% by weight skimmed milk powder were blocked at room temperature for 2 hours, the blocking solution was discarded, and the coated plate was air-dried;

[0067] 2) Adding samples: Add th...

Embodiment 3

[0077] According to the following steps, we tested the Aβ in the serum of the above 10 cases of healthy persons and 4 cases of AD patients 1-42 The antibody level was tested again, the concentration of the phosphate buffer solution in this example was 0.01mol / L, and the pH value was 7.2.

[0078] 1) Coating: 96-well strips are made of polystyrene plates with good adsorption properties, and the difference between wells, plates and batches is CV1-42 Protein fragment 10ng / ul solution 100ul, 4°C overnight, then discard the coating solution, wash the coated plate twice with PBS containing 0.1% TW-20 by volume, pat dry, add 1% bovine Serum albumin (BSA) and PBS with 1% by weight skimmed milk powder were blocked at room temperature for 2 hours, the blocking solution was discarded, and the coated plate was air-dried;

[0079] 2) Adding samples: add the serum to be tested from the human body to each hole of the coated strip, the amount of serum added is 100ul / well, the standard curve i...

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Abstract

The disclosed detection method for Abeta1-42 antibody comprises: covering solid holder by the Abeta1-42 protein fragment; adding the target sample into the holder to cultivate and clean the non-combined impurity; adding Abeta1-42 mono-clone antibody solution to cultivate and clean any the non-combined; detecting the content of combined component in the mixture. This invention has well accuracy and fit to quantitative detection in wide clinic application.

Description

1. Technical field [0001] The invention relates to the field of medicine and health, in particular to a method for detecting Aβ 1-42 Antibody methods and kits thereof. 2. Background technology [0002] At present, my country has entered an aging society, and Alzheimer's disease (AD) is a common and frequently-occurring disease of the elderly, and its incidence rate is as high as 2‰. AD is a primary degenerative disease of the central nervous system, with progressive decline in intelligence and cognitive function. Senile plaques (SP) and neurofibrillary tangles (NFT) are two major pathological changes of AD. Amyloid β (Aβ) is the main component of SP and participates in the formation of NFT. Aβ is a polypeptide containing 39-43 amino acids, which is produced by cleavage of its amyloid precursor protein (APP). The main component of SP in AD patients is Aβ 1-42 . At present, there is still a lack of laboratory-assisted diagnosis of AD clinically. [0003] In recent years,...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/547G01N21/78
Inventor 刘俊华
Owner 刘俊华
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