Method for quickly preparing drying agent array for protein crystallization

A protein crystallization and desiccant technology, applied in the field of protein crystallization, can solve the problems of time-consuming and labor-intensive silica gel desiccant, inability to realize popularization and application, and achieve the effect of improving production efficiency, improving success rate, and realizing popularization and application.

Inactive Publication Date: 2014-12-10
NORTHWESTERN POLYTECHNICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In order to overcome the time-consuming and labor-intensive preparation of silica gel desiccants of the required quality in the prior art, and the inability to realize popularization and application, the present invention provides a method for quickly preparing desiccant arrays, which can shorten the time for preparing desiccants and realize semi-automation. Save labor and improve production efficiency

Method used

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  • Method for quickly preparing drying agent array for protein crystallization

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1: Screening of lysozyme crystallization conditions.

[0024] 1. Preparation of desiccant array

[0025] Replace the ink in the inkjet printer with a cobalt chloride solution with a concentration of 100mg / ml, set the shape of the printed particles to be circular on the computer, and have a diameter of 0.5mm to form a 12×8 array with a particle spacing of 9mm. A solution of cobalt chloride was printed onto the glossy side of glossy paper, forming an array of solid cobalt chloride particles. Then stick the transparent sealing tape with glue on one side to the smooth surface of the glossy paper, tear off the tape from the glossy paper, and the array of solid desiccant particles adheres to the glued surface of the tape.

[0026] 2. Preparation of lysozyme crystallization solution

[0027] Lysozyme recrystallized six times (Japan Seikagaku Company, product number 100940) was dissolved in pH 7.00, 0.025mol / L HEPES-Na buffer to prepare a lysozyme solution with a conc...

Embodiment 2

[0031] Example 2: Screening of sweet protein crystallization conditions.

[0032] 1. Preparation of desiccant array

[0033] Use an electric pipette to pipette the mixed solution of 20 mg / ml cobalt chloride and 10 mg / ml aluminum chloride onto the smooth surface of the glossy paper to form a 12 × 8 array with a particle spacing of 9 mm and a droplet volume of 0.5 μl, dried, and then glued to the smooth surface of the glossy paper with a transparent sealing tape covered with glue on one side, and the tape was torn off from the glossy paper, and the array of solid desiccant particles adhered to the glued surface of the tape.

[0034] 2. Preparation of sweet protein crystallization solution

[0035] Sweet protein (Sigma, USA, product number T7638) was dissolved in pH 7.00, 0.025 mol / L HEPES-Na buffer solution to prepare a sweet protein solution with a concentration of 5 mg / ml. Use the automatic pipetting system to index Hampton company product number HR2-144 TM The protein crysta...

Embodiment 3

[0039] Example 3: Screening of lactalbumin crystallization conditions.

[0040] 1. Preparation of desiccant array

[0041] Use an automatic pipetting system to pipette the magnesium chloride solution with a concentration of 30 mg / ml onto the smooth surface of the glossy paper to form a 12×8 array with a particle spacing of 9 mm and a droplet volume of 0.3 μl. Adhesive transparent sealing tape sticks to the smooth surface of the glossy paper, and the tape is torn off from the glossy paper, and the array of solid desiccant particles adheres to the glued surface of the tape.

[0042] 2. Preparation of lactalbumin crystallization solution

[0043] Dissolve lactalbumin (Sigma, USA, product number L5385) in pH 7.00, 0.025mol / L HEPES-Na buffer to prepare a lactalbumin solution with a concentration of 10mg / ml. Use the automatic pipetting system to index Hampton company product number HR2-144 TM The protein crystallization reagents in the 96 crystallization kit were transferred to t...

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Abstract

The invention provides a method for quickly preparing a drying agent array for protein crystallization. The method comprises the following steps of: firstly, preparing a compound solution, and preparing a solid drying agent array on a scotch tape; secondly, preparing a single or a plurality of protein crystallization solutions according to experiment requirements; thirdly, sealing a crystal plate by using the scotch tape on which the solid drying agent array is pasted, and observing the result of crystallization; and finally, replacing the scotch tape, and repeating crystallization treatment under a closed condition until all the protein crystallization solutions are dried. By adopting the method, the success rate of protein crystallization is improved, the drying agent array can be quickly prepared, and the production efficiency is improved; and meanwhile, due to the high-consistency moisture absorption rate of compounds, the repeatability of crystallization is improved at the same time. The method can be used for realizing mass production for popularization and application.

Description

technical field [0001] The invention relates to a method for quickly preparing a desiccant array. The method can improve the preparation efficiency of the desiccant and is used to improve the success rate of protein crystallization, belonging to the field of protein crystallization. Background technique [0002] According to the statistics of the Structural Genome Project, the success rate of growing diffraction-quality protein crystals from very pure proteins is less than 20%. The process of obtaining protein crystals is one of the steps with the lowest success rate in the series of processes from the beginning of expression to obtaining the structure. Therefore, growing high-quality protein crystals is still one of the bottleneck problems in the field of structural biology. [0003] An important reason for the low success rate of protein crystallization is that there is no suitable crystallization agent to grow protein crystals. The most commonly used protein crystalliza...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K1/14C12N9/36C12N9/50C12N9/22C07K14/00
Inventor 尹大川鹿芹芹解旭卓闫二开陈达陈瑞卿刘永明
Owner NORTHWESTERN POLYTECHNICAL UNIV
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