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A method to improve the success rate of protein crystallization

A technology for protein crystallization and success rate, applied in peptide preparation methods, chemical instruments and methods, organic chemistry, etc., can solve problems such as poor adsorption capacity, difficult adsorption, large protein molecules, etc., to improve the success rate of crystallization and improve the shape Nucleation rate, effect of improving hydrophilicity

Active Publication Date: 2016-03-16
BEIJING ABACE BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since this kind of crystallization plate is a commercial product, the surface of the material used is smooth and has poor adsorption capacity, while the protein molecule is large and difficult to adsorb, which makes the protein solution rely on the heterogeneous nucleation of the wall to cause certain obstacles.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] 1. Preparation of surface treatment solution. Add potassium dichromate into distilled water, stir until potassium dichromate is completely dissolved, then slowly add concentrated sulfuric acid to the potassium dichromate solution and keep stirring until the solution is evenly mixed, and the mass ratio of each component in the obtained homogeneous solution is : Potassium dichromate: Distilled water: Concentrated sulfuric acid = 1: (2-5): (25-40).

[0017] 2. Handling of the crystal plate. The prepared solution is placed in a constant temperature water bath, and the temperature of the water bath is set at room temperature to 70°C. After the solution temperature is uniform, the crystallization plate is placed in the solution for 2 to 24 hours.

[0018] 3. Protein crystallization experiments. ProteinaseK, thaumatin, catalase, concanavalinA, α-chymotrypsinogenAI, ribonucleaseAIII, ribonucleaseAXII, subtilisinAVIII, papain, lactalbumin, HenFegg-whitelysozyme, Glucoseisomera...

experiment example 2

[0020] 1. Preparation of surface treatment solution. Add potassium permanganate into distilled water, stir until the potassium permanganate is completely dissolved, then slowly add concentrated sulfuric acid to the potassium permanganate solution and keep stirring until the solution is evenly mixed, the mass ratio of each component in the obtained uniform solution For: potassium permanganate: distilled water: chromic anhydride: concentrated sulfuric acid = 1: (1-5): (3-5): (5-25).

[0021] 2. Handling of the crystal plate. The prepared solution is placed in a constant temperature water bath, and the temperature of the water bath is set at room temperature to 70°C. After the solution temperature is uniform, the crystallization plate is placed in the solution for 2 to 24 hours.

[0022] 3. Protein crystallization experiments. ProteinaseK, thaumatin, catalase, concanavalinA, α-chymotrypsinogenAI, ribonucleaseAIII, ribonucleaseAXII, subtilisinAVIII, papain, lactalbumin, HenFegg-...

Embodiment 3

[0024] 1. Preparation of surface treatment solution. Add chromic anhydride into distilled water, stir until the chromic anhydride is completely dissolved, then add phosphoric acid to the chromic anhydride solution and stir, and finally add concentrated sulfuric acid to the solution slowly and keep stirring until the solution is evenly mixed, and the components in the obtained uniform solution The mass ratio is: chromic anhydride: distilled water: phosphoric acid: nitric acid: concentrated sulfuric acid = 1: (3-10): (10-40): (15-20): (60-120).

[0025] 2. Handling of the crystal plate. The prepared solution is placed in a constant temperature water bath, and the temperature of the water bath is set at room temperature to 70°C. After the solution temperature is uniform, the crystallization plate is placed in the solution for 30 minutes to 12 hours.

[0026] 3. Protein crystallization experiments. ProteinaseK, thaumatin, catalase, concanavalinA, α-chymotrypsinogenAI, ribonuclea...

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Abstract

The invention provides a method for improving success rate for crystallization of protein. The method comprises the following steps: uniformly mixing a solute, a solvent and water according to a certain sequence and proportion to prepare a solution with special components and proportion; placing the prepared solution in a constant temperature water bath; setting the temperature range from normal temperature to 70 DEG C; after the temperature of the solution is stabilized, soaking a crystallization plate in the prepared surface treatment solution; etching for a period of time to prepare different types of solutions for crystallization of protein; adding the prepared crystallization solution to the surface-treated and untreated crystallization plates; culturing in a constant temperature incubator; and observing the experimental result.

Description

technical field [0001] The invention relates to a method for improving the success rate of protein crystallization. Background technique [0002] The analysis of protein structure plays a very important role in explaining the physiological activities related to protein function. Although there are many methods for analyzing protein structure, the X-ray single crystal diffraction method for analyzing protein structure is still the most important, mature and widely used technique. And for this technique, protein crystals are the required medium. Therefore, the acquisition of protein crystals has become a key factor in the analysis of protein structures using X-ray single crystal diffraction technology. Convenient and efficient protein crystal growth technology has become the premise of using this technology to analyze protein structure. [0003] The heterogeneous nucleation method of adding a nucleating agent to promote nucleation can improve the nucleation rate of the prot...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K1/30
Inventor 尹大川何进郭云珠张辰艳刘永明曹慧玲崔超施建宇商澎
Owner BEIJING ABACE BIOTECH
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