Regulation and optimization apparatus for protein crystallization

Abstract

The invention relates to an apparatus of regulating and optimizing protein crystallization experiments by using a plurality of means. The apparatus comprises a protein solution pool, a precipitant solution pool, a quality sensor, a microscope, and a water circulation insulation cover. The apparatus uses modular combination to design a protein crystallization chamber, the protein solution pool and the precipitant solution pool are all independent modules, and distances of the two pools can be adjusted. The protein solution pool is located on a support, and quality change of the protein solution pool can be transmitted to the quality sensor with sensitivity higher than ten thousandth of a gram and then is quantitatively recorded. A baffle plate with adjustable hole size is mounted between the two solution pools, and a movable transparent glass block is arranged on top of the crystallization chamber so as to adjust a volume of a vapor phase. Meanwhile, the apparatus is provided with self light sources of a white light LED and a 280-nm ultraviolet light LED, thereby being capable of monitoring a growth status of the protein crystal in real time and identifying whether the crystal is the protein crystal. The apparatus can quantitatively and real-time monitor dynamical changes of a crystallization system caused by different regulation.

Description

Technical field: [0001] The present invention relates to a device for optimizing protein crystallization experiments using various regulation and control techniques. Background technique: [0002] An important prerequisite for using X-ray crystallography to study the structure of biological macromolecules such as proteins and further develop protein drugs is to make proteins form high-quality single crystals. However, due to too many factors affecting protein crystallization, crystallization has become the rate-limiting step in the entire process of protein structure and function research. [0003] Our study shows that the geometry of the crystallization chamber has a significant impact on protein crystallization kinetics and crystallization effectiveness. However, at present, laboratory crystallization uses crystal culture plates with fixed geometric dimensions, and some important parameters such as the distance from the protein solution to the precipitant solution, the vo...

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Problems solved by technology

However, at present, laboratory crystallization uses crystal culture plates with fixed geometric dimensions, and some important parameters such as the distance from the protein solution to the precipitant solution, the volume of the vapor phase, and the area of ​​the diffusion channel cannot be adjusted. culture plate

[0004] The patent "biological macromolecule crystallization device capable of automatic in-situ monitoring of vapor phase diffusion rate" (98203277.3) is an early invention of our research group. It can monitor the quality change of the protein solution through the mass sensor, and can also replace the precipitant solution, but cannot Simultaneously adjust the geometric parameters as described above

Invented "A Device for Cultivating Protein Crystals" (00233508.5) by piercing holes or inserting capillaries in the sealing film of commercial culture plates to change the vapor phase diffusion rate. Although it can be said that the diffusion channel area has been changed, it cannot be adjusted. Other geometric parameters do not have quantitative monitoring function

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