About HIC adsorption, Kads and Kdes

Hi!
I am totally new in CADET. However I was encouraged to learn CADET since we think that it can help us to see deeper in our chromatography process, hoping to perform a better separation for our target protein.

We have several questions as we are working with this platform to look at the Kads and Kdes of our sample using HIC.

  1. We’re wondering whether limitations are exist regarding protein concentration data which we input. The value of our concentration data (both target and impurities) is quite low (~0.02-2 mg/mL), and we’re afraid that too low concentration may not be proper to be used.
  2. Would it be possible for us to look at kads / kdes when our target is contained in a sample mixture, where the ‘impurities’ have similar properties to our target? (example: a pool of sample after undergoing digestion process, hence it contains initial substrate, intermediate products, protein target (around 60-70%), and nonspecific digested product). We’re asking since our team cannot acquire kads/kdes from the data we put in.
  3. Is there non ideal conditions (specifically on the sample properties) where it’s not probable to look at the kads/kdes?

Thank you in advance. We’re open to suggestions & corrections.