I tried to separate different proteins using Rp-Hplc.. My column C8-4.6x150mm..with pore size 5micron.. I could not separate this 2 proteins, they have same retention time.. Any suggest?
I would try a pore size of 300 Angstrom and as the stationary phase a C4 or a CN- ligand. Gradient water/acetonitrile with 0.1 TFA from 25 %-60% B (0-20 min). Temperature 35°C.