In order to study biomolecules, it is necessary to isolate them and purify them afterwards.

Purification of biomolecules allow scientists to shed light on biological processes by analyzing

its components separately. Aside from this, purification also assures that a sufficient amount of
the biomolecule being studied is obtained (Lehninger, Nelson & Cox, 2013). This is done to
assure the validity of the results and findings of the study.

With that being said, there are numerous techniques for separation and purification of
biomolecules. One of these techniques is chromatography. This process makes use of the
differential affinity between a mobile and a stationary phase. The mobile phase is usually a
liquid or gas that is permitted to flow over or flow through the stationary phase, which is a
column of spherical particles. A separation of the components of a protein mixture occurs once
the mixture is introduced to the mobile phase and allowed to migrate through the column. In
principle, the proteins with a higher attraction to the solid phase will migrate more slowly than
the proteins with affinity for the mobile phase (McMurry, 2011). Other examples of methods of
separation and purification of biomolecules are electrophoresis and ultracentrifugation. For the
purpose of this exercise, we will focus on chromatography.

Gel filtration or gel permeation or molecular sieve chromatography is a type of chromatography

that takes advantage of molecular size and shape differences, making it an effective way to
separate proteins with varying molecular weights. As discussed earlier, this is a column
chromatography where a bed of cross-linked gel particles, usually beads of one or two kinds of
polymers, are utilized as the stationary phase.Once the protein mixture is applied on the
column, the bed of porous beads will act as a semipermeable membrane and let only smaller
particles enter, whereas the larger particles are will be eluted or washed away by a solvent
(Campbell, Smith & Peters, 2006).

Some examples of the gel particles used are dextran and agarose, which are more commonly
known with their brand names Sephadex and Sepharose, respectively. Another gel used it Bio-
Gel, a polyacrylamide. For the purpose of this exercise, Sephadex, in the form of a dry powder,
is to be used. The goal of this exercise is to purify the crude egg albumin isolated in the previous
experiments and determine its molecular weight.