Ow. Reif et R. Freitag, CHARACTERIZATION AND APPLICATION OF STRONG ION-EXCHANGE MEMBRANE ADSORBERS AS STATIONARY PHASES IN HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY OF PROTEINS, Journal of chromatography, 654(1), 1993, pp. 29-41
Filtration membranes carrying strong cation- or anion-exchange groups
on their surface were evaluated for their potential as membrane adsorb
er stationary phases in the high-performance liquid chromatography of
proteins. The membranes are commercially available and can be obtained
inserted into ready-to-use filter holders. Owing to their thinness (1
70-290 mu m), the pressure drop of the membranes is extremely low. Flo
w-rates of up to 65 mi min(-1) per unit became thus possible. The low
pressure drop of a single membrane layer also permitted an effortless
scaling up, as a stack of several membranes or filter units could be u
sed, if necessary. Sample distribution, protein binding capacity, elut
ion conditions, separation efficiency and recovery were investigated a
s a function of the flow-rate. The time required for the separation of
certain protein mixtures could be reduced to less than 1 min. Appropr
iate conditions were defined for the separation of human serum and for
the isolation of subtilisin Carlsberg and beta-galactosidase from cel
l culture supernatants.