Bio-Rad BioLogic Signal Import Module (SIM) - Manual (Page 154)

Table 7-11
Scouting
To setup a scout experiment for method optimization.
Scouting is a procedure used to systematically optimize the purification of a specific target molecule (i.e.
protein). Molecules differ from one another in their charge, hydrophobicity, solubility, reactivity, substrate
specificity and in their intermolecular interactions. A purification protocol that is satisfactory for one type of
molecule may not work for a different molecule type. Several factors influence the quality of a purification
procedure. Some of these factors include: buffer composition (pH, ionic strength, co-solutes), elution type
(gradient slope and gradient duration), flow rate, column chemistry and sample composition. In principle,
each of these can be adjusted to produce the most efficient and effective purification strategy for a molecule.
In practice, only a few of these are generally tested due to time and cost considerations. By performing a
series of automated scout runs, the time and resources required for protocol optimization can be significantly
reduced.
Any method can be turned into a Scout Method using the Protocol Editor's Scouting Wizard. Methods used
in scout experiments can be copied from an existing method, created from scratch or loaded from the Bio-Rad
method templates. Before starting the Scouting Wizard, make sure the method setup and protocol are
correct and that the step parameters are set to the values desired for the first scout run.
Scouting Wizard Step 1
This dialog is used to select the type of scout to be carried ran.
• pH: Used in Buffer Blending mode to find the optimal pH for a chromatography method.
• Buffer A: Used to automatically run an experiment using a variety of load, wash or elution buffer
conditions through Inlet A.
• Buffer B: Used to automatically run an experiment using a variety of load, wash or elution buffer
conditions through Inlet B.
• %B: Used to systematical alter the salt composition of the buffer mobile phase during isocratic flow
and Load/Inject steps.
• Duration: Used to systematically vary the duration of isocratic flow, load/inject and linear gradient
steps.
Scouting Wizard - Step 1
Next > > Finish < < Back Cancel
pH
Type of Scout:
Buffer A
Buffer B
%B
Duration
%B (Gradient, Initial %B)
%B (Gradient, Final %B)
Flow Rate
Sample Name
Sample Volume
Column
Scouting
MODES OF OPERATION SYSTEM OPERATION
7-25