2013-15871

2016

Date Due: February 3,

Date Submitted:

February 3, 2016
EXPERIMENT NO. 4
PROTEIN CHARACTERIZATION BY GEL FILTRATION CHROMATOGRAPHY
Background of the experiment
Results and Discussion
In choosing the matrix, chemical stability, physical stability, and lack of adsorptive
properties were considered. It had perfect flow characteristics casted in bead so
column packing made easy. Pores within the beads had well-defined range of pore
sizes so that large molecules couldnt pass through some pores while smaller
molecules could. Choosing a gel was relevant on the type of the resin material since
it permits separation of different sizes due to having a variety of different ranges of
pore size in the beads.
Longer and wider column used in Gel Filtration Chromatography had a better
separation for high molecular weight polymers since more adsorbent particles are
passed by the protein bands, thus, a higher plate number. Sample added in a small
volume produced a better resolution due to diffusion and non-ideal flow effects,
thus, the volume containing proteins kept increasing.
In measuring the void volume, blue dextran was used due to its high molecular
weight. Blue dextran didnt enter the gels pores because of its large molecular
weight so it was eluted together with the void volume.
The ratio Ve/Vo was used since it only depends entirely on the size of the molecules
and is independent on the concentration of protein and the size of the column.