Purification Of Protein By Two-Dimensional HPLC

RPC-HILIC-SCX: This sequence was used to obtain polypeptides of extremely high purity from crude pituitary extracts for correction of mistaken sequence data in the literature (actually, a SCX-RPC-HILIC sequence would have been more convenient). Ref: K.F. Faull et al., Neuropeptides 32 (1998) 339.

PROTEINS : 
Ion-exchange and hydrophobic interaction chromatography (HIC) are useful for most protein applications, particularly if protein tertiary structure and function is to be preserved (not a concern in proteomics). The capacity and resolving power of SEC is much less than for these modes, making it useful only in special cases. The organic solvents used for RPC and HILIC also make them useful only for special cases. Small proteins tend to work well with RPC while proteins < 30 KDa and membrane proteins frequently work well with HILIC.

RPC-WCX : This combination worked well for this small protein. It would have been more convenient to use a WCX-RPC sequence.

Column Name : PolyCAT A™ Columns

Compound Name : Polypeptides