Method for increasing biological macromolecule crystal mass

A technology of biological macromolecules and crystal quality, which is applied in the preparation methods of peptides, crystal growth, chemical instruments and methods, etc., to achieve the effect of more crystal surface morphology, improved resolution, and regular crystal surface morphology

Inactive Publication Date: 2011-04-13
NANTONG CAMBRIDGE OLEIN +1
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Problems solved by technology

[0004] In order to solve the problem of poor quality of biological macromolecular crystals crystallized in the prior art, the present invention provides a method for improving the quality of biological macromolecular crystals, using a certain frequency and amplitude Mechanically vibrate the biological macromolecule crystallization solution system, and strictly control the temperature of the vibration environment, which can improve the quality of protein crystals

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  • Method for increasing biological macromolecule crystal mass

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[0013] Step 1: Put the vibrating table into the biochemical incubator, and the temperature of the vibrating table is controlled by the biochemical incubator. The model of the vibration table is RT-40D, the vibration direction of the vibration table is vertical, the vibration frequency is 4-200Hz, and the amplitude is 2×10 -4 ~5mm; the model of the biochemical incubator is LRH-250, the temperature control range of the biochemical incubator is 0-60°C, the temperature control accuracy is ±0.1°C, and the temperature fluctuation is ±0.5°C.

[0014] The second step: the lysozyme whose product number is 100940 of Japan Seikagaku Company through six times of recrystallization is dissolved in pH 4.60, 0.1mol / L sodium acetate buffer to make its concentration 20mg / ml, and use 0.22μm filter to remove impurities. Use the same buffer solution to prepare NaCl crystallization solution with a concentration of 80mg / ml. Use the hanging drop method and use a 40-hole crystallization plate for bio...

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Abstract

The invention discloses a method for increasing biological macromolecule crystal mass, comprising the steps: a biological macromolecule crystallizing solution system is prepared and then fixed on a vibration table with the vibration frequency of 0.1-3000Hz and the vibration amplitude of 2*10<-4>-10mm, and the vibration table has the environmental temperature control accuracy of 0.1 to -0.1DEG C and the temperature fluctuation of 0.5 to -0.5 DEG C; the biological macromolecule crystallization is carried out at the constant temperature of 0-60 DEG C in the vibration; after crystallization, the crystal is recovered. As the vibration frequency of 4-3000Hz and the vibration amplitude of 2*10<-4>-1.0mm are adopted, mechanical vibration is carried out on a protein crystallization plate, and the temperature is strictly controlled within the range of 0-60 DEG C, the increased amplitude of the resolution of the biological macromolecule crystal can be 0.3-1.6, and the appearance of the crystal surface is more regular.

Description

technical field [0001] The invention relates to a method for improving the quality of biological macromolecular crystals, in particular to a method for improving the quality of biological macromolecular crystals through mechanical vibration. Background technique [0002] The study of the structure and function of biological macromolecules is a hot issue in the frontier field of biology. Structural information of biological macromolecules is very helpful for the design and development of new drugs. X-ray diffraction is currently the most reliable method for testing and determining the three-dimensional structure of biological macromolecules, and the implementation of this method requires high-quality crystals of biological macromolecules. The success rate of growing from high-purity proteins to diffraction-quality protein crystals is less than 20%. Therefore, obtaining high-quality biomacromolecular crystals is still a bottleneck problem. [0003] Many factors affect the c...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K1/14C30B30/06
Inventor 尹大川鹿芹芹王玺凯卢慧甍杨鹏飞商澎
Owner NANTONG CAMBRIDGE OLEIN
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