HPLC Analysis of MAb on Proteomix^® SCX-NP5 Comparing Different Buffer Systems and pH Gradients

column

Proteomix SCX-NP5, 25 cm x 4.6 mm I.D., 5 μm particles (Z777161)

mobile phase

Mobile phase A: (A) 20 mM MES, pH 6.0; (B) A + 1 M NaCl, pH 6.0Mobile phase B: (A) 2.4 mM Tris, 1.5 mM imidazole, 11.6 mM piperazine, pH 6.0; (B) A, pH 10.5

gradient

Gradient A: 7% B held for 2 min; to 14% B in 20 min, Gradient B: 0 to 30% B in 2 min; to 55% B in 30 min

flow rate

0.8 mL/min

detector

UV, 280 nm

injection

5 μL

sample

10 mg/mL