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Acidified albumin and preparation method thereof, and application of acidified albumin in inhibiting beta-amyloid protein aggregation

A technology for amyloid and serum albumin, applied in the field of biomedicine, can solve the problems of in vitro modification of proteins, and achieve the effect of reducing cytotoxicity, reducing toxicity, and increasing the number of negative charges

Inactive Publication Date: 2016-02-10
TIANJIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] There have always been great difficulties in modifying proteins in vitro

Method used

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  • Acidified albumin and preparation method thereof, and application of acidified albumin in inhibiting beta-amyloid protein aggregation
  • Acidified albumin and preparation method thereof, and application of acidified albumin in inhibiting beta-amyloid protein aggregation
  • Acidified albumin and preparation method thereof, and application of acidified albumin in inhibiting beta-amyloid protein aggregation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1: Synthesis and characterization of acidified serum albumin with a molar ratio of diethylene glycol anhydride to serum albumin of 10

[0037]Weigh 100 mg of serum albumin and dissolve it in 20 mL of deionized water to obtain a serum albumin solution with a concentration of 5 mg / mL. Weigh 1.75 mg of diethylene glycol anhydride powder and add it into the serum albumin solution, while adding dropwise 1 mol / L sodium hydroxide solution to maintain the pH of the reaction solution at 6.0. The above mixture was stirred at 170 rpm and room temperature for 0.5 h. After the reaction, the free diglycolic anhydride in the system was removed through a SephadexG-25 gel filtration chromatographic column, and the obtained product (acidified serum albumin 1) was freeze-dried and stored.

[0038] Weigh 2 mg each of serum albumin and acidified serum albumin 1, dissolve them in 2 ml of buffer solution with pH 3.0, 4.0, 5.0, 6.0, 7.0, 8.0, and 9.0 respectively; ) were used to meas...

Embodiment 2

[0045] Example 2: Synthesis and Characterization of Acidified Serum Albumin with Diglycolic Anhydride and Serum Albumin Molar Ratio of 100

[0046] Weigh 200 mg of serum albumin and dissolve it in 20 mL of deionized water to obtain a serum albumin solution with a concentration of 10 mg / mL. Weigh 35.07 mg of diglycolic anhydride powder and add it into the serum albumin solution, while adding dropwise 1 mol / L sodium hydroxide solution to maintain the pH of the reaction solution at 6.5. The above mixture was stirred at 170 rpm and room temperature for 2 h. After the reaction, the free diglycolic anhydride in the system was removed through a SephadexG-25 gel filtration chromatographic column, and the obtained product (acidified serum albumin 2) was freeze-dried and stored.

[0047] Weigh 2 mg each of serum albumin and acidified serum albumin 2, dissolve them in 2 ml of buffer solution with pH 3.0, 4.0, 5.0, 6.0, 7.0, 8.0, and 9.0, and pass a potentiometric analyzer (Malvern Instr...

Embodiment 3

[0054] Example 3: Synthesis and characterization of acidified serum albumin with a molar ratio of diethylene glycol anhydride to serum albumin of 400

[0055] Weigh 400 mg of serum albumin and dissolve it in 20 mL of deionized water to obtain a serum albumin solution with a concentration of 20 mg / mL. Weigh 280.53 mg of diglycolic anhydride powder and add it to the serum albumin solution, while adding 1 mol / L sodium hydroxide solution dropwise to maintain the pH of the reaction solution at 7.0. The above mixture was stirred at 170 rpm and room temperature for 5 h. After the reaction, the free diglycolic anhydride in the system was removed through a SephadexG-25 gel filtration chromatographic column, and the obtained product (acidified serum albumin 3) was freeze-dried and stored.

[0056] Weigh 2 mg each of serum albumin and acidified serum albumin 3, dissolve them in 2 ml of buffer solution with pH 3.0, 4.0, 5.0, 6.0, 7.0, 8.0, and 9.0 respectively; ) were used to measure th...

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Abstract

The invention relates to an acidified albumin and a preparation method thereof, and an application of the acidified albumin in inhibiting beta-amyloid protein aggregation. According to the invention, diglycolic anhydride is modified on the amino group on the surface of serum albumin, such that a product with isoelectric point of 3.8-4.3 and with diglycolic anhydride average modification number between 3 and 14.5 is obtained. The product is the acidified albumin. Diglycolic anhydride powder is added into a solution of serum albumin dissolved in deionized water and with a concentration of 1-20mg / mL; a sodium hydroxide solution is dropped in at a same time, such that the pH value of the reaction liquid is stably maintained at 6.0-7.0; a stirring reaction is carried out for 0.5-5h under room temperature; and free diglycolic anhydride is removed, such that acidified serum albumin with a stable molecular structure is obtained. According to the acidified serum albumin provided by the invention, serum albumin surface negative charge amount is improved, such that the interaction of serum albumin and Abeta is promoted, and thus Abeta aggregation is inhibited, and beta-amyloid protein aggregate cytotoxicity is reduced. The acidified albumin is used in medicines used for treating Alzheimer's disease and beta-amyloid protein aggregation-related diseases.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to an acidified albumin, a preparation method and an application in inhibiting the aggregation of beta-amyloid protein. Background technique [0002] Alzheimer's disease (AD) is the most common neurodegenerative disease, and its pathological manifestations are clinical symptoms such as cognitive and memory dysfunction caused by extracellular neural plaques and intracellular neural tangles. , can lead to death in severe cases. As of 2011, there were 35 million AD patients worldwide, and the incidence of AD increased significantly with age. As the problem of aging society continues to intensify, AD has become the fifth leading cause of death among the elderly, and it has brought a heavy economic burden to families and society. [0003] A large number of studies have shown that the main pathological feature of AD is the deposition of amyloid plaques outside of cerebra...

Claims

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Application Information

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IPC IPC(8): C07K14/765C07K1/113A61K38/38A61P25/28
Inventor 孙彦谢宝龙史清洪董晓燕
Owner TIANJIN UNIV
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