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Protein crystallization apparatus, method of protein crystallization, protein crystallizing agent and process for preparing the same

A technology of protein crystallization and crystallization agent, which is applied in the field of protein crystallization agent and its preparation, and can solve the problems of complex operation, high price of automatic equipment, mixed pollution, etc.

Inactive Publication Date: 2006-11-15
MITSUBISHI RAYON CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] However, in the above-mentioned protein crystallization apparatus, movement, mixing (contamination) of protein-containing samples and / or crystallization agents may occur between regions of the microarray.
Therefore, there is a possibility that the conditions for crystallization are inconsistent with the concentration and type of the crystallization agent contained in the gel-like substance in the preliminary zone.
[0008] In addition, the use of such a device requires manual work to supply a protein-containing sample to the crystallization agent holding part, and the operation becomes complicated.
However, automatic devices for automatically supplying protein-containing samples are expensive

Method used

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  • Protein crystallization apparatus, method of protein crystallization, protein crystallizing agent and process for preparing the same
  • Protein crystallization apparatus, method of protein crystallization, protein crystallizing agent and process for preparing the same
  • Protein crystallization apparatus, method of protein crystallization, protein crystallizing agent and process for preparing the same

Examples

Experimental program
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Effect test

Embodiment 1

[0185] Weighing 0.48g of sodium chloride as precipitating agent is put into 10ml volumetric flask, constant volume with 0.1M-citric acid buffer (pH4.0), prepares 1.2M-NaCl aqueous solution (this solution is designated as a liquid). In addition, 5 g of N, N'-methylenebisacrylamide (hereinafter abbreviated as MBAAm) and 5 g of deionized water were added to 90 g of a 50% aqueous solution of acrylamide (manufactured by Mitsubishi Rayon Co., Ltd.) to prepare a 50% monomer solution. (This solution is designated as liquid b). Furthermore, 2,2'-azobis[2-(2-imidazolin-2-yl)propane] dihydrochloride (manufactured by Wako Pure Chemical Industries, VA-044) as a water-soluble polymerization initiator was prepared in 10 % aqueous solution (this solution is defined as liquid c). Inject 840 μl of solution a and 160 μl of solution b into a 2 ml sample tube and mix to prepare an aqueous protein crystallization agent solution (1.0M-NaCl, pH 4.0, monomer concentration 8%), then add 10 μl of sol...

Embodiment 2

[0187] In addition to changing the concentration of liquid a in Example 1 to 2.4M, the others are prepared according to the same procedure as the crystallization agent aqueous solution (2.0M-NaCl, pH4.0, monomer concentration 8%), and then follow the same procedure Steps to obtain a transparent hydrogel. Also, the formation of lysozyme crystals was confirmed by the same method. The results are shown in Table-1.

Embodiment 3

[0188] Except changing the concentration of liquid a in Example 1 to 3.6M, other all prepare crystallization agent aqueous solution (3.0M-NaCl, pH4.0, monomer concentration 8%) according to the same procedure, then follow the same procedure Steps to obtain a transparent hydrogel. Also, the formation of lysozyme crystals was confirmed by the same method. The results are shown in Table-1.

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Abstract

A protein crystallization apparatus with which protein crystallization experiments and crystallization condition screening can be carried out rapidly and economically with high reliability; and a protein crystallizing agent with which the operation of protein crystallization can be carried out in a simple manner. In particular, the protein crystallization apparatus is characterized by including protein crystallization microarray (18) having two or more crystallizing agent holding parts (16) adapted to retain a protein crystallizing agent and, superimposed on the protein crystallization microarray (18), plate (24) wherein the plate (24) has crystallization compartments (32) corresponding to the above crystallizing agent holding parts (16) and adapted to be charged with a protein-containing sample and has recessed parts (34) interposed between crystallization compartments (32). The protein crystallizing agent is characterized in that the crystallizing agent is uniformly held in a gel as a result of gelation of a solution containing a protein precipitating agent and an unsaturated monomer.

Description

technical field [0001] The invention relates to a device and method for protein crystallization or screening of crystallization conditions, and a protein crystallization agent for precipitating protein crystals from protein-containing samples and a preparation method thereof. Background technique [0002] In recent years, research called structural genomics is underway, which attempts to elucidate the function of a gene encoding a protein by elucidating the relationship between the spatial structure of the protein and the function of the protein. [0003] The analysis of the spatial structure of proteins usually starts with screening the crystallization conditions of the protein to be analyzed. Then, crystallization is carried out under optimum crystallization conditions. Analysis of the spatial structure of the protein is carried out by subjecting the obtained crystals to X-ray structure analysis. Among them, in the process of screening protein crystallization conditions,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K1/30C30B29/58
Inventor 田中熏渡边信久竹内浩史井关隆幸涩谷望西岛千晴
Owner MITSUBISHI RAYON CO LTD