Memory improving mixture separated from blood plasma and preparation method and application of memory improving mixture separated from blood plasma

A technology for enhancing memory and mixture, applied in the field of plasma isolates to achieve the effect of improving cognition, delaying disease progression and enhancing memory

Active Publication Date: 2016-11-09
北京豪思生物科技股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the specific components of these substances and the methods of separation and acquisition have not been reported yet.

Method used

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  • Memory improving mixture separated from blood plasma and preparation method and application of memory improving mixture separated from blood plasma
  • Memory improving mixture separated from blood plasma and preparation method and application of memory improving mixture separated from blood plasma
  • Memory improving mixture separated from blood plasma and preparation method and application of memory improving mixture separated from blood plasma

Examples

Experimental program
Comparison scheme
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preparation example Construction

[0056] A method for preparing the above-mentioned mixture separated from plasma, the mixture is obtained by performing a series of treatments on the plasma, and the treatment steps include plasma collection, density gradient centrifugation, dialysis, primary concentration, removal of high peak protein, anion Exchange and secondary concentration steps. details as follows:

[0057] Various culture mediums and commonly used reagents used in the present invention are all prepared by conventional methods. If the molecular biology operations involved in the embodiments are not specified as specific test conditions and methods, please refer to editors such as SambrookJ, Science Press, 2002, Molecular Cloning Experiment Guide (Third Edition) or the instruction manual of the corresponding product.

[0058] In the plasma collection step, the blood is collected using an anticoagulant tube, and the supernatant is collected by centrifugation to obtain plasma; , 1100g, 1200g, 1300g, 1400g...

Embodiment 1

[0077] Embodiment 1: the preparation of mixture

[0078] (1) Collect blood from adult mice and collect plasma

[0079] Take 100 adult mice of C75BL / 6 strain (this mouse can also be replaced by other strains of experimental animals, such as: mice of BALB / cA strain, mice of DBA / 2 strain, SD rats, Wistar rats ) anesthetized with 20vol% carbon dioxide (that is, a gas with a carbon dioxide content of 20vol%), and then use a blood collection needle to bleed blood from the mouse's eyes into a blood collection tube (the blood collection tube is an anticoagulant tube), and shake the blood collection tube while bleeding to prevent blood coagulation . Put the blood collection tube into the centrifuge, set the rotating speed to 1000g, and centrifuge for 30 minutes. The supernatant is then carefully aspirated with a pipette, which is the collected plasma.

[0080] (2) isolate the cocktail component i.e. the mixture of the present invention from mouse plasma

[0081] (a) Density gradien...

Embodiment 2

[0093] Example 2: Identification of cocktail components prepared in Example 1 by SDS-PAGE denaturing gel electrophoresis

[0094] Take 2 microliters from the above-mentioned cocktail components, and measure its absorbance under ultraviolet 280nm, so as to calculate the protein concentration of the cocktail components. Take a certain volume of cocktail components and mix them with 1 microliter of 5× protein sample buffer (purchased from Beijing Lamboride Biotechnology Co., Ltd., Cat. No. D621), which is the sample to be electrophoresed. There are 10 micrograms in the sample of protein. The electrophoresis sample was heated up to 100°C, heated for 20 minutes to denature the protein, then the sample was immediately placed on ice, and after waiting for 5 minutes, the SDS-PAGE denatured reducing gel was run (the preparation method of the SDS denatured reducing gel is as follows: 30( w / v) % acrylamide Acr-Bis (purchased from GE Healthcare) to take 1.3ml, 1.5M Tris-hydrochloric acid...

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Abstract

The invention discloses a memory improving mixture separated from blood plasma and a preparation method and application of the memory improving mixture separated from the blood plasma. The mixture comprises various proteins and various micromolecules, SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) of the mixture comprises at least 30 bands, and the bands with molecular weights larger than 48kD include seven evidently visible bands with the molecular weights being 56kD, 68kD, 77kD, 115kD, 175kD, 250kD and 289kD sequentially. The preparation method of the mixture includes steps of blood plasma collection, density gradient centrifugation, dialysis, primary concentration, high-abundance protein removal, anion exchange and secondary concentration. The mixture prepared according to the method not only has a memory improving function but also can be used for preventing, improving and treating Alzheimer's disease.

Description

technical field [0001] The present invention relates to a plasma isolate, in particular to a mixture separated from plasma for enhancing memory and its preparation method and application. The mixture can be used to prevent, improve and treat Alzheimer's disease, and It has the effect of enhancing memory. Background technique [0002] Alzheimer's syndrome, also known as Alzheimer's disease, is characterized by a gradual loss of memory and cognitive abilities in patients. Patients die within 3 to 9 years of diagnosis, accounting for 50% to 56% of clinical deaths. The current understanding of the cause of the disease is the accumulation of abnormally folded beta-amyloid and tau proteins in the brain that lead to Alzheimer's disease. [0003] The drugs currently on the market for Alzheimer's disease are mainly western medicines, which are characterized by long-term adherence to taking them, but have little curative effect, can only alleviate some symptoms, and cannot control t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/16A61P25/28
CPCA61K9/0002A61K35/16
Inventor 崔文宏
Owner 北京豪思生物科技股份有限公司
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