Module 5 : Analysis of biomolecules

Lecture 32 : Gel Filtration chromatography

As per relationship given in Eq 32.3, three different type of analytes are possible:

1. Analyte with Kd=0, or Ve=Vo , these analytes will be completely excluded from the column.

2. Analyte with Kd=1 or Ve=Vo+Vi , these analytes will be completely in the pore of the column.   

3. Analyte with Kd>1, in this situation analyte will adsorb to the column matrix.

Figure 32.3: A typical Gel Filtration Chromatogram.

Choice of matrix for gel filtration chromatography-The choice of the column depends on the range of molecular weight and the pressure limit of the operating equipment. A list of popular gel filtration column matrix with the fractionation range are given in Table 32.1.