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Process for producing crystalline nucleus and method of screening crystallization conditions

A manufacturing method and screening method technology, applied in solution crystallization, chemical instruments and methods, single crystal growth, etc., can solve problems such as insufficient crystallization, difficult protein crystallization, etc., and achieve rapid and definite results

Inactive Publication Date: 2005-10-05
CHUANGJING CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, sufficient crystallization is not possible using this method, especially protein crystallization is difficult

Method used

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  • Process for producing crystalline nucleus and method of screening crystallization conditions
  • Process for producing crystalline nucleus and method of screening crystallization conditions
  • Process for producing crystalline nucleus and method of screening crystallization conditions

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0070]Put DAST (4-dimethylamino-N-methyl-4'-N-styrylpyridinium tosylate) 3.5g and methanol 200ml together with the rotor into a polytetrafluoroethylene container 6 with a volume of 200ml, It was heated to 55.0° C. in the constant temperature water tank 8 with an initial temperature of 27.0° C. over 2 hours, and dissolved while stirring it. After about 5 hours, it was confirmed that DAST was dissolved, and the solution 7 was divided into 3 parts, and used as a culture solution. At this point, remove the rotor. After the solution was prepared for 16 hours, it was heated at 55°C for 10 hours, followed by a 3°C drop per hour to bring the temperature down to 23°C. And drop 0.1°C per hour, bringing the temperature down to 21.4°C. In this state, the pulse energy is 250μJ / pulse (2×10 9 watts) and a repetition rate of 1 kHz to irradiate the femtosecond laser for 2 minutes. Thereafter, no crystallization was confirmed for 1 hour, and 10 hours later, crystallization was observed with...

Embodiment 2

[0072] Egg white lysozyme was used as a target for crystal nucleation. This solution was adjusted to pH 4.5 by adding 0.467 g of sodium acetate trihydrate to 50 ml of distilled water, adding acetic acid thereto, and then adding 1.25 g of sodium chloride and 1.25 g of egg white lysozyme. The aforementioned sample solution 7 adjusted to room temperature was placed in a 100 ml polytetrafluoroethylene container 6, and kept in a constant temperature water bath 8 at 40° C. for 24 hours to completely dissolve it. After that, it was cooled to 25° C. over 5 hours, and impurities were removed with a membrane filter. 2 ml of this solution and 3 ml of perfluorocarbon (Florinate) were respectively added to ten glass bottles 6 with screw caps having a diameter of 18 mm. These glass bottles 6 were kept at 25°C and left still in a constant temperature water tank 8, and the temperature of the solution was lowered to 15°C over 20 hours. The saturation point of this sample solution (lysozyme s...

reference example 1

[0076] use figure 2 The shown setup investigates the shock wave generated by the explosive phenomenon using a pulsed laser. In this device, a pulsed laser irradiation device is mounted on an erecting microscope. As shown in the figure, the erect microscope 11 is provided with a stage 28 on which an object to be observed is placed, a condenser lens 29 and an objective lens (100 times, aperture number 1.25) 26, and a microchip 27 is placed on the stage 28. In addition, a light source lamp 13 is disposed below the condenser lens 29 at the lower portion of the erect microscope 11 , and a CCD camera 12 for detecting the light is disposed above the microscope 11 . In addition, a pulsed laser irradiation device 21 is arranged outside the erect microscope 11, and the laser light 22 is irradiated in the erect microscope 11 through the 1 / 2 wavelength plate 23 and the polarizer 24, and the optical path is bent at a right angle by the dichromatic mirror 25, and irradiated on the object....

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Abstract

The present invention relates to a process for producing high-quality crystals of protein or organic substances easily and efficiently. A solution of protein or an organic substance is prepared and then is cooled slowly to be supersaturated to a low degree. This supersaturated solution is irradiated with a femtosecond laser 10. A local explosion phenomenon occurs at the focal point of the laser and thereby a crystalline nucleus is generated. A high-quality crystal is obtained when a crystal is grown on the crystalline nucleus over a long period of time. The femtosecond laser to be used herein can be a titanium:sapphire laser having a wavelength of 800 nm, a duration of 120 fs, a frequency of 1 kHz, and an output of 400 mW.

Description

technical field [0001] The present invention relates to a method for producing crystallization nuclei and a screening method for crystallization conditions. Background technique [0002] With the development of post-genome research, the structural analysis of proteins is becoming a top priority. For this, the protein needs to be crystallized. In addition, organic crystals are promising as next-generation device materials, and their high-quality crystal manufacturing technology is particularly required. Generally, in order to precipitate crystals from a solution, it is necessary to increase the degree of supersaturation by solvent evaporation, temperature change, or the like. However, substances with large molecular weights such as organic substances and proteins cannot be crystallized unless their supersaturation is very large. In addition, in a solution in which a very large degree of supersaturation is formed as described above, once crystals are generated, there may be...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C30B7/00
CPCC30B7/00Y10S117/904C30B29/58
Inventor 佐佐木孝友森勇介吉村政志安达宏昭增原宏细川阳一郎高野和文
Owner CHUANGJING CO LTD
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