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)