Bio-Rad Nuvia™ cPrime™ Media - Manual (Page 12)

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1. Load feed or eluate from previous step directly without
dilution onto the Nuvia cPrime media column. To elute, use an
increasing pH gradient. If satisfactory elution and recovery are
achieved, refine and/or make a step gradient to complete the
step (range pH 4–8, depending on protein).
2. If elution is not satisfactory after step 1, run a salt gradient to
disrupt electrostatic or hydrophobic interactions that may be
preventing elution or broadening the peak. Use the pH where
there was best elution (from step 1). The direction of this salt
gradient (increasing or decreasing) can be easily assessed
and will depend on the relative contributions of ionic vs.
hydrophobic interactions involved in binding.
3. If elution is still unsatisfactory after step 2 of this process,
consider using a modifier such as propylene glycol, urea, or
arginine to disrupt any remaining interactions. Other modifiers
may also be used; in some cases changing to another salt
may also be required.
For further assistance or to discuss method development,
contact Bio-Rad chromatography technical support group
at 1-510-741-6563.