Purification of biomolecules

Intro:
Gel Permeation Chromatography (GPC) is used in this experiment. This process allows molecules to be separated on the basis of their size using a column which contains a special separation matrix. An important consequence of this structure is that other molecules can enter these beads and it is possible to tailor the properties of these gel beads so that only relatively small molecules can enter while larger molecules are excluded.
Changes made to the protocol
# Liquid-liquid extraction was not carried out following the gel permeation chromatography in order to extract cholesterol.
# Detection of reducing sugars was not carried out using Benedict’s reagent. Another type of reagent was used that turned red/pink in the presence of reducing sugars.
# 1ml fractions were collected from the GPC instead of 2ml.



Discussion:

From the result we understand the small molecules such as glucose in this case is able to enter the beads and hence become partially trapped and passes in between.
The overall understanding we get is that the large molecules went through the beads
more faster in comparison with small molecules. Using this known fact protein and the glucose became separated and purified.

From the figures we understand the protein is seen in fraction 3, it is at its peak
concentration at fraction 5 & 4. It overlaps with the glucose in fractions 5&6. We can see further on in the fraction that the concentration is decreases.