Bio-Rad Foresight™ Chromatography Columns, Prepacked - Manual (Page 14)

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The time interval used for sample addition dictates
the number of time points. At the end of the longest
incubation time the liquid in all wells is separated
from the media, preferably through centrifugation.
The experiment uses a fixed phase ratio. The
amount of bound solute is calculated using the mass
balance presented in Eq. 2.
• A time between 20 and 60 minutes is usually
enough for an uptake study. However, the actual
value depends on the molecular properties,
experimental conditions (e.g., ionic strength), and
binding kinetics
• The contact time needed to achieve a target
load can de determined using uptake curve data,
for a given phase ratio and initial total solute
concentration
2.6 Contact Time
The incubation or contact time refers to the time the
sample containing the solute is in contact with the
chromatographic media. Contact time affects the
amount of solute bound to the media (capacity) for
a fixed phase ratio and initial solute concentration.
Relatively long incubation times lead to equilibrium
capacities. Conversely, relatively shorter times lead to
capacities located along the operating line Figure 1A.