Bio-Rad Foresight™ Chromatography Columns, Prepacked - Manual (Page 20)

Technical Support: 1-800-424-6723 • www.bio-rad.com 15
Csalt (M) …
C
0
(mg/mL) 1 2 3 … 10 11 12
C
1
A
C
2
B
C
3
C
C
4
D
C
5
E
C
6
F
C
7
G
C
8
H (A)
Csalt
1
Csalt
4
C
0
(mg/mL) C
1
C
2
C
3
… C
10
C
11
C
12
Salt (M) 1 2 3 … 10 11 12
A
B
C
D
E
F
G
H
(B)
Csalt
1
Csalt
2
Fig. 3. Schematic representation of the plate
configuration for isotherm experiments using a fixed
phase ratio, and variable initial solute concentration
(C
0
). A, configuration suitable for an 8-channel pipet;
B, configuration suitable for a 12-channel pipet. This
example suggests performing three replicates for each salt
concentration.
3.2.3 Binding Kinetics
This type of study is useful for defining
the contact time of the batch adsorption
experiment. For a fixed phase ratio and initial
solute concentration, the contact time dictates
how much solute will bind to the media. If
there is enough sample and time, it is best to
perform the binding kinetic study using two
sufficiently distinct initial solute concentrations.
This is especially useful if no isotherm data
are available. The longest incubation time for
uptake experiments is usually 60 minutes,
meaning equilibrium is not reached, especially
for macromolecules such as proteins and
antibodies. Figure 4 shows the loading sequence
(Figure 4A) and media plate configuration (Figure
4B) for the uptake experiments.
A