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A beta 1-10 specific binding peptide and screening method thereof

A specific, peptide-binding technology for biological applications

Inactive Publication Date: 2013-01-30
陈贵海
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

There are few drugs currently approved for clinical treatment of AD, only acetylcholinesterase inhibitors such as donepezil and N-methyl-D-aspartate receptor antagonists such as memantine, but these drugs cannot fundamentally reverse AD AD condition, its treatment is still at a symptomatic level

Method used

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  • A beta 1-10 specific binding peptide and screening method thereof
  • A beta 1-10 specific binding peptide and screening method thereof
  • A beta 1-10 specific binding peptide and screening method thereof

Examples

Experimental program
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Effect test

Embodiment Construction

[0040] Materials and methods used in this embodiment are as follows

[0041] 1.1 Main Consumables

[0042] ELISA polystyrene microwell experimental reaction plate

[0043] short peptide Aβ 1-10 and Biotin-Aβ 1-10 Synthesized by Xi'an Huachen Company

[0044] Streptavidin (Streptavidin, SA) was purchased from Sigma, USA

[0045] Biotin (NHS-LC-Biotin) was purchased from Sigma, USA

[0046] 1.2 Main reagents

[0047] 1.2.1 Culture medium and formula

[0048] See Table 3. Adjust pH to 7.0, add distilled water to constant volume, T 121°C, P 0.103×10 6 Pa, autoclave for 30min.

[0049]Table 3 medium and formula

[0050]

[0051]

[0052] 1.2.2 Solution

[0053] PEG / NaCl: 20% PEG, 150mM NaCl

[0054] TEA: 0.1M

[0055] Tris-HCl: 1mM Tris, adjust pH to 7.4 with HCl

[0056] TBS: 50mM Tris-HCl and 150mM NaCl add water to volume, 0.103×10 6 Pa autoclave for 30min

[0057] IPTG / X-gal: 1.25g IPTG and 1g X-gal dissolved in 25ml dimethylformamide

[0058] PBS: first ...

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Abstract

The invention discloses an A beta 1-10 specific binding peptide and a screening method thereof. A beta 1-10 on A beta molecules, which comprises segments influencing mutual aggregation of the A beta molecules is used as a target molecule. A short peptide capable of being specifically bond with the special fragment is screened by using a phage peptide library display technology. Specific binding A beta is observed in vitro, and the capability of automatically generating plaques by the A beta is inhibited. The screening method comprises the following steps of: first, synthetizing A beta 1-10 comprising an EFRH sequence, and carrying out biotinylation on part of the A beta 1-10; then, with biotinylated A beta 1-10 as the target molecule, screening out phages with high specificity and strong affinity by adopting a liquid affinity capture method in the phage display technology; detecting the affinity dynamics relationship between the screened phages and the target molecule with a biomolecule interaction analyzer; observing whether the specific phages can prevent the A beta molecules from mutually aggregating to form the plaques under a transmission electron microscope; selecting monoclones for the specific phages at random, and sending to a sequence test company to test the sequence subjected to PCR (Polymerase Chain Reaction) amplification; and synthetizing corresponding short peptides according to test results.

Description

[technical field] [0001] The invention relates to the field of biotechnology, in particular to an Aβ 1-10 Specific binding peptides and screening methods thereof. [Background technique] [0002] With the prolongation of life expectancy and the decrease of birth rate year by year, the problem of aging in my country is becoming more and more serious. The high prevalence of dementia in the elderly population has become a serious social problem. Alzheimer's disease (AD) is the most common cause of dementia in the elderly. At present, there are few drugs approved for clinical treatment of AD, only acetylcholinesterase inhibitors such as donepezil and N-methyl-D-aspartate receptor antagonists such as memantine, but these drugs cannot fundamentally reverse AD The treatment of AD is still at a symptomatic level. At present, the generally accepted hypothesis is that the increase of Aβ load in the brain is the initiating factor of the pathological process of AD. Therefore, substa...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K7/08C07K1/00
Inventor 陈贵海王芳许文华周先岭杨启纲曹磊
Owner 陈贵海