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Methods

Along with the progress of structural analysis, analysis difficult factor has come to be analyzed. This is important in a case, such as the purification (such as oxygen-free-Sugumiru Law and automated crystallization system) (various, such as use and construction of mass culture-based expression systems) and crystallization, is to establish a high-quality sample preparation method. There we are focused.

Protein Crystallization System (PXS)

Dispensing to the crystallization plate, storage in the incubator, is a system that performs fully automatic drop of photography. Drop photo taken according to the schedule it is possible to browse from all over the world through a web browser.

Large scale expression and purification of target proteins

By the protein, since there is compatibility with the expression system, such as E. coli, yeast, insect, mammalian cells, and offers a wide variety of expression systems. It also takes advantage of cell-free expression system in the joint research.
In addition, (a few g from a few mg) a large amount of purified protein crystallization is required, it is important to prepare a large amount of starting material for this purpose. Currently, we have established an experimental setup that can be repeated cultivation of approximately 100 liters every day.

Development of new methods for crystallization

There is a good thing when a crystallized in an oxygen-free conditions. First, it will be able to crystallize the reaction intermediate stop the oxidation-reduction reaction. In addition, you do not have to loss of protein it is possible to prevent the formation of oxide films. In addition, it is difficult to handle even can awaited crystal and can oxide film, but it can be difficult to apply the X-ray in the crystal, in the oxygen-free conditions you can prevent this.
Also, after mixing the protein solution and the crystallization solution, it is possible to immediately see and observe the Shometsu precipitation, we have developed a method which can be estimated to close to crystallization conditions conditions.
In addition, we are developing a method to improve the quality of the crystal by soaking in a multi-stage anti-freezing agent.

Equipment for physico-chemical analysis

The purified protein as a physical and chemical analysis of the means,

  • Circular dichroism dispersion meter [JASCO, J-820]
  • Static light scattering [Wyatt, DAWN HELEOS-II]
    And dynamic light scattering [Wyatt, DynaPro NanoStar]
  • Biacore [GE, Biacore T200]
  • MALDI TOF-MS [Bruker, autoflex-M2]
    Peptide sequencer [AB, Procise]
  • Dual-wavelength fluorescence scanner [LI-COR, Odyssey]
Such as, you can use the device in house.