Bio-Rad Foresight™ Chromatography Columns, Prepacked - Manual (Page 8)

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Normally, V
0
is much larger than the latter
two liquid volumes, hence the assumption
that V = V
0
is a reasonable approximation.
Implementing this assumption from Eq.1 leads
to the following expression for the concentration
of adsorbed solute (binding capacity):
( )
0
0
ads
amount bound
volume of adsorbent
V
q C C
V
= − =
According to Eq. 2, the amount of bound material
can be calculated using the feed concentration (C
0
),
the flowthrough (unbound) concentration (C), the
volume of sample added (V
0
), and the volume of
adsorbent present in a given well (V
ads
).
A mass balance for multiple steps in a given well
(e.g., flowthrough (FT) collection, washing, elution,
and regeneration) is presented in Eq. 3a-3d:
loaded bound unbound
m m m = +
loaded 0 0
m C V =
bound wash wash elution elution regen regen
m V C V C V C = + +
unbound FT FT
m V C =
Where m is the mass in a given stage, C represents
the solute liquid phase concentration in the various
stages [i.e., loading (C
0
), flowthrough collection (C
FT
),
washing (C
wash
), elution (C
elution
), and regeneration
(C
regen
)], and V represents the liquid volume used in a
given step [i.e., loading (V
0
), washing (V
wash
), elution
(V
elution
), and regeneration (V
regen
)]. The amount of
bound protein can be estimated using either Eq. 3a
or Eq. 3c; the amount of bound solute can be more
easily estimated using Eq. 3a due to the simplicity of
the experimental measurements required to obtain its
inputs.
Eq. 3b
Eq. 3c
Eq. 3d
Eq. 2
Eq. 3a