Aim:

To pack a gel filtration column and determining the void volume of the packed column

Introduction:

Gel filtration chromatography is also known as gel permeation, molecular sieve, and size exclusion chromatography. The molecules are separated based on their size. The column matrix is made up of small spherical porous beads. The smaller molecules can enter the pores present in the beads while the molecules larger than the maximum pore size of the bead are completely excluded. The access to the pores is determined by both the shape and the molecular weight of the molecules; the separation, therefore, is based on the ability of the molecules to enter the porous beads. Properties of some of the routinely used gel filtration matrices are shown in table 22.1.

Table 22.1: Gel filtration matrices commercially available for separation in the range from 100 Da to subcellular particles