Question

how would SEC (size-exclusive chromatography) enhance (or not) for protein purification and why? What advantages or disadvantages? What could

be done in the future for greater purification?

Answer 1

Solution

• Size exclusion chromatography is the process of separation of mixture of protein sample according to there size shape and molecular weight in gel column.
• it is helpful to enhance the purification of proteins having different sizes because it contains solid phase of network of smaller sized beads which are helpful to retard the smaller proteins inside them.
• larger proteins will elute first and medium sized second and smaller are inside solid phase.

Advantages

• Low chances of sample loss
• Requires less time for separation and analysis
• Biological activity is preserved while during process.
• Narrow band obtained with good sensitivity.
• The flow rate of separation can be set.

Disadvantages

• The limited number of peak can be resolved within short time on gel column.
• Dust or other particles change of interfere on the peak value detection.
• Large sized column needs more eluent for separation.
• While separation proteolysis can happen and it is a major problem.
• It has low resolution than the other chromatographic techniques.

what could be done for in a future for greater purification.

• To prevent proteolysis of proteins some strategies can adapt for gretaer purification.
• If pH of liquid phase is maintained then there will no chances of change in biological activities of protein complex sample.
• If it is combined with moleculer weight and charges then it will be a very powerfull technique for future protein seperation perspective.