Bio-Rad Foresight™ Chromatography Columns, Prepacked - Manual (Page 18)

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3.2.2 Adsorption Isotherm
An adsorption isotherm can be determined using
a ladder of initial solute concentrations and a
fixed phase ratio. This approach is suitable for
plates loaded with a constant volume of media.
Figure 3 presents plausible ways of configuring
the Foresight™ chromatography media filter
plate for an isotherm experiment done in
triplicate. The presented examines the effect
of salt concentration on the isotherm. The two
alternatives depicted differ on whether an
8-(Figure 3A) or 12-(Figure 3B) channel pipet is
available. To streamline the workflow, load the
solutions in a 96-deep well plate, transferring
them to the media plate using the multichannel
pipet. The time required to reach equilibrium may
range from 1–6 hours of incubation, depending
on target protein and buffer conditions. The
incubation should be done under mixing. To
minimize evaporation, the media plate should be
covered with a sealing film or a sealing mat.