Gel Filtration Chromatograph Technique

Gel Filtration Chromatograph Technique

Gel Filtration Chromatograph Technique

Question:

Discuss about the Gel filtration chromatograph technique.

Answer:

Introduction And Background

The principal method that is used for the separation and purification of the biomolecules in biochemical laboratories is chromatography. In chromatographic separation, partition occurs in between two phases one is mobile phase and another one is stationary phase. Gel filtration chromatography or size exclusion chromatography is a type of chromatography (Murray, Granner, Mayes and Rodwell 2009). It is used to separate protein molecules based on stokes radius. Stoke radius is the measured diameter of the sphere occupied by the protein when they tumble in the solution. Stoke radius can also be defined as the function of molecular mass and shape (Murray, Granner, Mayes and Rodwell 2009).  As protein with large stroke radius are too large to enter inside the porous bed (stationary phase) and are thus eroded away (mobile phase) where as protein with smaller stoke radius enter the indentation of the porous bed and remain there until they are drifted back by the mobile phase. The protein thus separated emerge from the gel filtration column in the descending order of their measured stroke radii (Murray, Granner, Mayes and Rodwell 2009; Nelson, Lehninger and Cox 2008).

The following report gives a detailed analysis of concept behind the gel filtration chromatography along with the potential advantages and disadvantages associated with it. The report further illustrates the mechanism of gel filtration chromatography via diagrammatic representations and two video links. After illustrating the concept, the report tends to shed light on the practical application of this biochemical procedure in research along with different porous beds with their size limits, which are commonly employed as stationary phase. At the end, the report will provide a detailed breakup of the formula used to calculate the protein size in gel filtration chromatography.

What’s It Used For

Gel filtration chromatography is the simplest technique used for the separation of the complex component by according  to their molecule size only, measuring by how dynamic they penetrate the pore of the stationary phase, this  chromatography is also know a size exclusion chromatography or molecular exclusion chromatography. This type of method  permits to purified polypeptide from other polypeptides based to their size by moving through the gel porous into the column. Gel filtration is application use to analyze the separating of molecules size such as protein, polysaccharides -and nucleic acids by separating and removing large proteins and complexes.