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A device for studying protein crystallization using periodically pulsating magnetic fields

A protein crystallization, pulsating magnetic field technology, applied in the direction of material magnetic variables, etc., to achieve the effect of stable temperature, improving the success rate of crystallization, and promoting gas phase diffusion

Inactive Publication Date: 2015-11-18
NORTHWESTERN POLYTECHNICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the effect of continuously changing dynamic magnetic field on protein crystallization has not been reported in the literature.

Method used

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  • A device for studying protein crystallization using periodically pulsating magnetic fields
  • A device for studying protein crystallization using periodically pulsating magnetic fields
  • A device for studying protein crystallization using periodically pulsating magnetic fields

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1: Periodically pulsating magnetic field promotes gas phase diffusion

[0028] Step 1: Set up the periodic pulsating magnetic field generator. Set at 20°C, rotate at 300rpm, and adjust the magnetic field strength to 0.78T.

[0029] The second step: preparing protein crystallization solution. The Hen's Egg-WhiteLysozyme freeze-dried powder (Hen's Egg-WhiteLysozyme) from Seikagaku Company's article number E05801 was dissolved in HAC-NaAC buffer (0.1M, pH4.60) to prepare lysozyme solutions with different concentrations, respectively 40, 60, 80mg / ml. Dissolve NaCl in this buffer solution to prepare a precipitant solution with a concentration of 60 mg / ml. Different concentrations of lysozyme solutions are mixed with the same concentration of precipitant in equal proportions by volume to prepare a protein crystallization solution.

[0030] The third step: crystallization. The crystallization method is the commonly used hanging drop method, and the "cup-shaped dis...

Embodiment 2

[0037] Example 2: Periodic pulsating magnetic field improves crystallization success rate

[0038] Step 1: Set up the periodic pulsating magnetic field generator. Set at 20°C, rotate at 300rpm, and adjust the magnetic field strength to 0.78T.

[0039]The second step: preparing protein crystallization solution. Lysozyme solution (20 mg / ml) was prepared by dissolving egg white lysozyme freeze-dried powder of Seikagaku Company with product number E05801 in HAC-NaAC buffer (0.1 M, pH 4.60). NaCl was dissolved in this buffer to prepare a precipitant solution (60 mg / ml). The lysozyme solution and the precipitating agent are mixed uniformly in equal proportions by volume to obtain a protein crystallization solution.

[0040] The third step: crystallization. The crystallization method is the MicroBatch method, and the "cup-shaped dish + arc-shaped support" petri dish is applied.

[0041] One side of the holder is sealed with crystallization tape to firmly adhere the cup to the ho...

Embodiment 3

[0046] Example 3: The periodic pulsating magnetic field makes the plane orientation of the crystal, and the orientation rate is different

[0047] Step 1: Set up the periodic pulsating magnetic field generator. Set at 4°C, rotate at 300rpm, and adjust the magnetic field strength to 0.85T.

[0048] The second step: preparing protein crystallization solution. Lysozyme solution (25mg / ml) was prepared by dissolving egg white lysozyme freeze-dried powder of Seikagaku Company with product number E05801 in HAC-NaAC buffer (0.1M, pH4.60). NaCl was dissolved in this buffer to prepare a precipitant solution (60 mg / ml). The lysozyme solution and the precipitating agent are mixed uniformly in equal proportions by volume to obtain a protein crystallization solution.

[0049] The third step: crystallization. The crystallization method is an air-free Batch method, and an injection-type curved support petri dish is used.

[0050] The 2 sides of the rack are sealed with crystallization t...

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Abstract

The invention provides a device for the research on protein crystallization by periodic pulsating field. The device comprises a container, two rotary discs, a motor and sample support tables and is characterized in that the interior temperature of the container is controlled to 0-60 DEG C, the rotary discs are disposed in the container and can be driven by the motor to synchronously rotate around a rotary shaft, a permanent magnet is disposed at one end of one rotary disc, far away from the rotary shaft, and samples are fixed in rotational projection planes of the rotary discs respectively through the sample support tables. The device is stable in temperature, allows for dynamically varying magnetic field and stable, non-vibrating and non-interference conditions for samples, and also allows for single-factor influencing of periodic pulsating field.

Description

technical field [0001] The invention relates to a periodic pulsating magnetic field generator. Background technique [0002] In the post-genome era, the study of protein structure and function is an inevitable trend. X-ray single crystal diffraction technology, as the most important technology, has resolved more than 88.0% of the protein structures in the protein structure database, and the growth of high-quality protein single crystals has always been a bottleneck restricting this technology. Therefore, people have been devoting themselves to the study of protein crystallization methodology, hoping to improve the success rate of crystallization condition screening and obtain high-quality protein single crystals through the development of various methods, effectively solving the bottleneck problem in this technology. Many methods have been researched and developed, and many successful experiences have been accumulated. [0003] Protein crystallization under magnetic field ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/72
Inventor 尹大川解旭卓樊艳婷曹慧玲
Owner NORTHWESTERN POLYTECHNICAL UNIV