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Protein aggregation inhibitor

A protein and inhibitor technology, applied in the field of protein aggregation inhibitors, can solve problems such as poor heat resistance, aggregation, and inability to fully inhibit protein aggregation, and achieve the effect of inhibiting aggregation and inhibiting activity reduction

Inactive Publication Date: 2018-01-02
OSAKA ORGANIC CHEM INDS +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, for example, proteins such as egg yolks and egg whites of eggs generally have poor resistance to heat and have the property of easily agglomerating due to heating. cohesion

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Add 3-[(3-acylamidopropyl)dimethylammonium]propane-1-sulfonate 3.925g, water 15 g, 45 g of methanol, and 0.2187 g of 2-(dodecyltrithiocarbonate)-2-methylpropionic acid were heated up to 70° C. and stirred while introducing nitrogen gas into the flask. Thereafter, 0.1947 g of azobisisobutyronitrile as a polymerization initiator was added to the flask with stirring, and the monomer components in the flask were polymerized for 6 hours to obtain a sulfobetaine polymer solution.

[0058] After the content in the flask was cooled to room temperature, it was taken out from the flask, and the weight average molecular weight of the obtained sulfobetaine polymer was analyzed by gel permeation chromatography (hereinafter referred to as GPC) using a device [manufactured by Tosoh Corporation, HLC8220GPC] As a result of measurement, the weight average molecular weight (in terms of polystyrene) of the obtained polymer was 5,500. In addition, when measuring the weight average molecula...

experiment example 1

[0070] Using the aqueous solution obtained in Comparative Example 1, 2 mL of this aqueous solution was mixed with 100 μL of Thioflavin T solution (PBS 16 μg / mL), and the mixed solution obtained above was measured using a model: T3516 manufactured by Sigma Aldrich (liquid temperature: about 25° C. ) the fluorescence intensity at an excitation wavelength of 440 nm and an emission wavelength of 480 nm, and obtain the basic fluorescence intensity based on the following formula.

[0071] [Basic fluorescence intensity] = [fluorescence intensity at emission wavelength] ÷ [fluorescence intensity at excitation wavelength]

[0072] Next, using the solution obtained in Example 1 to which lysozyme (PBS) was added, 2 mL of this solution was mixed with 100 μL of Thioflavin T solution (PBS 16 μg / mL), and the obtained mixed solution was used (liquid temperature: about 25° C.) When the fluorescence intensity was obtained in the same manner as above, and the ratio of the fluorescence intensity ...

experiment example 2

[0087] Lysozyme (PBS) was added at a ratio of 3 mg per 1 mL of the protein aggregation inhibitor obtained in Example 1. The obtained solution was observed visually, but aggregation of lysozyme was not confirmed.

[0088] Next, the solution obtained above was heated at a temperature of 90° C. for 30 minutes, and then cooled to room temperature. The cooled solution was observed visually, but aggregation of lysozyme was not confirmed.

[0089] Next, the time-dependent change in absorbance at a wavelength of 600 nm of the solution after heating at a temperature of 90° C. for 30 minutes was measured using a spectrophotometer [Shimadzu Corporation, model: UVPC1600]. As a result, the absorbance of the solution when heated at 90° C. for 30 minutes and cooled to room temperature was 0.85, and the absorbance of the solution after 6 minutes from cooling to room temperature was 0.8.

[0090] On the other hand, an aqueous solution of lysozyme was prepared by adding 3 mg of lysozyme (PBS)...

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Abstract

The present invention provides a protein aggregation inhibitor which can suppress protein aggregation, even if the protein is heated, and can suppress decreases in protein activity, even if the protein is further heated, the protein aggregation inhibitor characterized by comprising a sulfobetaine polymer resulting from polymerizing a monomer component comprising a sulfobetaine monomer representedby formula (I). (In the formula, R1 represents a hydrogen atom or a methyl group, R2 represents an alkylene group having 1 to 4 carbon atoms, R3 represents an alkyl group having 1 to 4 carbon atoms, R4 represents an alkylene group having 1 to 4 carbon atoms, and X represents an -NH- group or an -O- group.)

Description

technical field [0001] The present invention relates to protein aggregation inhibitors. The protein aggregation inhibitor of the present invention can effectively prevent protein aggregation even when the protein is heated, so it is expected to be used for various requirements such as enzyme preservatives, antibody drugs, and amyloid aggregation inhibitors in the body to prevent protein aggregation. The purpose of cohesion. Background technique [0002] As a protein aggregation inhibitor capable of preventing protein aggregation under acidic conditions, a protein containing a compound selected from polyoxyethylene distyrenated phenyl ether, polyoxyethylene myristyl ether, and polyoxyethylene (10) octylphenyl ether has been proposed. A protein aggregation inhibitor of at least one nonionic surfactant in ether (for example, refer to Patent Document 1). [0003] Since the aforementioned protein aggregation inhibitor uses a surfactant, it is possible to suppress protein aggreg...

Claims

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Application Information

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IPC IPC(8): C07K1/113A23J3/00C08F20/38C08F20/60C12N9/96
CPCA23J3/00C07K1/113C08F20/38C12N9/96C08F20/60C12N9/2462A23L15/30A23V2002/00
Inventor 松村和明R·罗宾古川刚前原贤太郎猿渡欣幸
Owner OSAKA ORGANIC CHEM INDS
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