Software Manual

6 | Creating a Method
180 | NGC Chromatography Systems and ChromLab Software
Scouting on Parameters within a Method
Scouting is a procedure used to systematically locate parameters that most
impact peak resolution and to optimize on these parameters to achieve your
protein purification goals (for example, high purity, yield, stability, or activity).
Molecules differ from one another in their charge, hydrophobicity, solubility,
reactivity, and substrate specificity, and in their intermolecular interactions. A
purification protocol that is satisfactory for one type of molecule might not work for
a different molecule type. Several factors influence the quality of separation in a
purification procedure. These factors include buffer composition (pH, ionic strength,
cosolutes), 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.
Using ChromLab’s scouting wizard you can easily generate a series of methods that
scout on a parameter within certain sections of the method.
Methods used in scout
experiments can be generated from an existing method, created from scratch, or
loaded from the Bio-Rad method templates.
Scouting methods can be saved using the Save As option. After you create a
scouting series of methods, changing the method outline will invalidate the scouting
series and you will be pr
ompted to scout again and save the method as a new
scouting method.