Separation of two closely related serine protease analogues using HPCE/MECC with SDS in a dynamic mode

HPCE was evaluated for the identification of different serine protease analogues. This work represents a separation of two serine proteases, Savinase (SAV) (Novo Nordisk A/S, Bagsvaerd, Denmark) and a Savinase analogue, both consisting of 269 amino acids and having identical isoelectric points. The only difference between the two protease variants is that a methionine is substituted with a serine, M222S. The separation was only made feasible by the HPCE/micellar electrokinetic capillary chromatography (MECC) technique using sodium dodecyl sulfate (SDS) dynamically as the surfactant. This paper shows that it is possible to employ HPCE/MECC to separate Savinase and a Savinase variant with no alteration in net charge.