Helpful questions

Target molecule

1. Acquaint yourself with the properties of the target molecule, e.g. size, conformation, stability
range, isoelectric point, etc.!
Molecular weight 464 Kda, homotetramer, between 37-75 degree Celsius,ip 4.61. This
product is stable for at least one year when stored at -0C.
2. What is the effect of pH on the protein charge (in relation to the isoelectric point)?
pH below their pI, proteins carry a net positive charge; above their pI they carry a net
negative charge.
3. How is the enzymatic activity influenced by the surrounding conditions? TEMPERATURE AND
SURROUNDING Ph. Like at above 75 degree.

Separation

4. What are the basic mechanisms involved in the separation? ATPS Mechanism
5. What are important influencing factors? Type of polymer/salt using, Ph
6. Draw a flowsheet that contains the essential steps needed to perform the respective unit
operation. Set up an overall mass balance and all relevant component balances.
7. Which auxiliary substances are involved? Inform yourself about their properties, availability
and prices.
8. At some points you will probably need to make assumptions because reliable information
might be difficult to obtain (at least without experiments). However, your assumptions
should be reasonable and justified (mention and explain them in your presentation/report).

Technical realization

What kind of equipment could be suitable?

Are there alternatives and, if yes, which aspects could be considered when making a choice?

What are the advantages/disadvantages of batch processes and continuous processes?

How is cleaning performed?

Does the process require frequent maintenance and what does it involve? Is it rather a high or low
throughput operation? Does it cause a high pressure drop?

What about the feasibility of scale-up, is it easy or difficult? How can it be done?

Does product inactivation or degradation occur? If yes, why and how? Can it be influenced?

Optimization and robustness

For which step(s) in the RIPP scheme is the unit operation applicable? Is there a possibility to
combine several steps in one (process integration)? By which means can the process be intensified
or improved? Do you see possibilities for e.g. heat integration to make the overall process more
efficient? Feed material, purity requirements and even products can vary or change. How flexible is
the process concerning such variations? What deviations from your theoretical calculations would
you expect when putting the process into practice?

Environmental