Which of the following are important characteristics to consider when designing

Question

Which of the following are important characteristics to consider when designing a protein purification strategy from a total eukaryotic cell lysate? a. The size of the protein. b. The cellular location of the protein. c. The biological activity of the protein. d. Two of the above e. All of the above. When using gel filtration (or size exclusion chromatography) which proteins, large or small elute off the column first? A cation exchange column is BEST suited to separate proteins that are negatively charged. True False

Explanation / Answer

1. Two of the above

The protein purification strategy involves biological activity, physics chemical property, protein size and binding affinity.

The location of protein (cytoplasm or within organelles) is considered only during protein extraction.

2. In gel filtration, the column is filled with beads which also has interstitial spaces. The smaller molecules enter the beads and take more time to elute. The larger molecules will escape through the interstitial spaces and will be eluted early.

3. False

The cation exchanger will have negatively charged resin which attracts positively charged particles. Hence it can be used to separate positively charged proteins.

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