美迪西--蛋白质晶体结构测定

蛋白质晶体结构测定
> 结晶条件筛选
> 蛋白质共结晶及小分子浸泡
> 三维结构解析：Se-MAD法和分子置换

美迪西各种结晶体图：

美迪西结晶方法：
1、Multiple constructs into multiple vector
• N- and C- terminal truncation
• Protein mutation of surface residue
• Remove of flexiable regions

2、Expression & Purification of Protein
• Expression & solubility Test
(Co-expression with chapaton or ligand)
• Pilot purification and optimization
(AC, IEX, HIC, SEC ,co-purification with ligand,
buffer optimization)
• Scale-up production (10-50 mg)
• Protein concentration (2-20 mg/ml)

3、QC of target protein
• Mass spectroscopy
• SDS-PAGE
• HPLC (Size exclusion column behavior

4、Crystallization trays set-up
• Around published condition
(5 mg compound)
• Kits screen (10-20 mg)

5、Optimization
• pH and pricipitatant
• Additive screen
• Seeding
• Redesign constructs

6、Diffraction-quality crystal