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.
Eriksen
J
J
Novo Nordisk Research Institute, Bagsvaerd, Denmark.
Holm
K A
KA
D001407:Bacillus / Q000201:enzymology; D002021:Buffers; D002197:Capillary Action; D002845:Chromatography / Q000295:instrumentation / Q000379:methods; D019075:Electrophoresis, Capillary / Q000295:instrumentation / Q000379:methods; D005285:Fermentation; D007202:Indicators and Reagents; D007527:Isoenzymes / Q000737:chemistry / Q000302:isolation & purification; D012697:Serine Endopeptidases / Q000737:chemistry / Q000302:isolation & purification*; D012967:Sodium Dodecyl Sulfate
D002021:Buffers; D007202:Indicators and Reagents; D007527:Isoenzymes; D012967:Sodium Dodecyl Sulfate; D012697:Serine Endopeptidases; C032923:microbial serine proteinases