The effects of pH and ionic strength (salt concentration) on transmission of single protein human serum albumin (HSA) and human immunoglobulins (HIgG) through a 100 kDa molecular weight cut-off (MWCO) polyethersulfone membrane have been studied using a pulsed sample injection technique. Experimental results obtained clearly demonstrate the effectiveness of changing solution pH and ionic strength optimisation for selective transmission of HSA or HIgG. These also demonstrate the usefulness of pulsed injection experimental technique for the optimization of ultrafiltration processes. A pulse input carrier phase ultrafiltration (CPUF) was employed to fractionate a binary protein mixture of HSA and HIgG at low NaCl concentration, showing that it is feasible to separate the binary protein at optimised conditions.