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Support having protein immobilized thereon and method of producing the same

A manufacturing method and support technology, applied in chemical instruments and methods, carrier binding/immobilizing peptides, specific peptides, etc., can solve problems such as loss of function, activity, inability to immobilize proteins, and low content rate

Inactive Publication Date: 2010-01-20
JSR CORPORATIOON
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, depending on the molecular type of the protein, a sufficient amount of protein may not be immobilized due to the small content of amino acids having amino groups or the small number of amino groups present on the surface of the three-dimensional structure of the protein that is likely to come into contact with the support. Case
In addition, for proteins that have been immobilized on a support, the amino groups that play an important role in the expression of protein functions and activities are consumed in binding to the support, resulting in loss of function and activity.

Method used

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  • Support having protein immobilized thereon and method of producing the same
  • Support having protein immobilized thereon and method of producing the same
  • Support having protein immobilized thereon and method of producing the same

Examples

Experimental program
Comparison scheme
Effect test

Synthetic example 1

[0052] 2.1. Synthesis Example 1 (Synthesis of Immobilization Support (Magnetic Particles) Having Carboxyl Group)

[0053] 2 parts by mass of 75% di(3,5,5-trimethylhexanoyl) peroxide solution ("PEROYL 355-75(S)" manufactured by NOF) was mixed with 1% sodium lauryl sulfate aqueous solution for 20 Parts by mass, finely emulsified with an ultrasonic dispersant. This was placed in a reactor containing 13 parts by mass of polystyrene particles with a particle diameter of 0.77 μm and 41 parts by mass of water, and stirred at 25° C. for 12 hours. Using another container, 96 parts by mass of styrene and 4 parts by mass of divinylbenzene were emulsified with 400 parts by mass of a 0.1% aqueous solution of sodium lauryl sulfate, and the resulting liquid was put into the above-mentioned reactor and stirred at 40°C. After 2 hours, the temperature was raised to 75°C, and the polymerization was carried out for 8 hours. After cooling to room temperature, the resultant was further washed wit...

Synthetic example 2

[0059] 2.2. Synthesis Example 2 (Synthesis of Immobilization Support (Magnetic Particles) Having Epoxy Group)

[0060] In Synthesis Example 1, the same steps as in Synthesis Example 1 were carried out except that 13.5 g of GMA and 1.5 g of TMP were used instead of 13.5 g of cyclohexyl methacrylate and 1.5 g of methacrylic acid. This gave particles having epoxy groups (hereinafter referred to as "particles B").

Synthetic example 3

[0061] 2.3. Synthesis Example 3 (Synthesis of Immobilization Support (Magnetic Particles) Having Toluenesulfonyl Group)

[0062] 5 g of particle B obtained by freeze-drying was put into a 1 L separable flask, 60 ml of 1 mol / L sulfuric acid was put therein, and the mixture was stirred at 60° C. for 6 hours. Next, the particles in the above-mentioned detachable flask were separated using magnetic force, and then washed repeatedly with distilled water.

[0063] In summary, magnetic particles having 2,3-dihydroxypropyl groups were obtained. 1.0 g of dry particles obtained by freeze-drying the particles were dispersed in 8 ml of pyridine, and then 0.2 g of p-toluenesulfonyl chloride was added, followed by stirring at room temperature for 2 hours. After the reaction, the particles were separated by magnetic force, washed four times with acetone, and then washed four times with distilled water to obtain magnetic particles in which 2,3-dihydroxypropyl was tosylated (hereinafter refer...

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PUM

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Abstract

A method of producing a support having a protein immobilized thereon which comprises immobilizing a protein having a tag sequence containing a sequence consisting of three or more consecutive basic amino acids on a support.

Description

technical field [0001] The present invention relates to a method for producing a protein-immobilized support capable of efficiently immobilizing a protein on the immobilization support, and to a protein-immobilized support on which a protein is immobilized. Background technique [0002] Regarding the support on which the protein is immobilized, the protein is used as a probe in order to purify or detect biological substances and chemical substances. For example, supports of protein A and protein G on which proteins having affinity with antibody molecules have been immobilized are used for affinity purification of antibodies. In addition, the support on which the antibody is immobilized is used as a diagnostic reagent for detecting and quantifying the antigen by antigen-antibody reaction. [0003] When a protein-immobilized support is used for such purification or detection, it is generally necessary to increase the amount of protein immobilized in order to improve the purif...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543G01N33/553C07K17/00
CPCC07K17/06G01N33/54353G01N33/543
Inventor 片寄聪福田哲夫村田充弘
Owner JSR CORPORATIOON