Final answer:
Protein separation in SEC is affected by column size, mobile phase composition, sample concentration, and flow rate. These need to be optimized for effective separation and resolution.
Step-by-step explanation:
Several factors can influence protein separation in SEC:
- The column size determines the range of molecular weights that can be effectively separated, with larger columns typically able to separate larger molecules.
- The composition of the mobile phase can affect how proteins interact with the column material and therefore influence separation.
- Sample concentration should be optimal to prevent overloading the column, which can lead to band broadening and poor separation.
- The flow rate affects how long proteins interact with the stationary phase and can impact resolution and separation efficiency.
Optimizing these factors can lead to improved separation and resolution of proteins during SEC, facilitating accurate identification and analysis.