Compare and contrast the different techniques used in separating biomolecules.

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Compare and contrast the different techniques used in separating biomolecules

Aim

The aim of protein purification is to remove or isolate one particular protein from a solution containing a mixture of other molecules

In this assignment, I will assess the effectiveness of different techniques used to separate bio molecules (proteins and lipids) by comparing these techniques and exploring the best technique for separating proteins products. I will also discuss the application of biomolecule separation techniques in medicine, industry and for economical reasons.

Introduction

Protein bioseparation refers to the recovery and purification of protein products from various biological materials and is important in food, pharmaceutical, and biotechnological industry. Protein based products need to be purified before they can be used.

Bio molecules produced in cell culture (whether in bacteria, yeast, or other cells need to be separated from potentially toxic components of culture. Column Chromatography and electrophoresis are some of the most used techniques for separating and purifying these substances.

Protein based products include

. food and nutritional products

2. pharmaceutical products

3. industrial catalysts

4. diaganostic products

biomolecular separation is based on properties such as size charge and function.

An ideal protein bioseparation process must combine high productivity with high selectivity of separationand must be feasible at mild operating conditions.

Proteins have disulphide, these are broken before separation.

SEPARATION OF PROTEINS

Column Chromatography of proteins

Colunm chromotography relies on the distribution of components do be separated in between two different phases, a mobile phase and a stationary phase. The mobile phase is used to carry the components involved through the stationary phase. The staionary phase is supplied through a packed bed in the column. There is a wide variety of chromatographic techniques; these are used to separate protein mixtures on the basis of molecular properties such as size, shape and weight or certain binding affinities. The three chromatographic techniques used to separate proteins are; Gel filtration, ion exchange and affinity chromatography. In such chromatographic methods, the protein mixture is dissolved in a liquid known as the mobile phase. During the stationary phase, protein molecules separate from each other because of their different distribution between the two phases. This technique is used mainly in small labaratories as it yields a productivity of purified protein but at high resolution.
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Gel filtration chromatography

Gel filtration chromatography separates proteins on the basis of there size and shape using a column packed with gelatinous polymer. Molecules that are larger than gel pores are excluded. Molecules that are smaller than the gel pores diffuse throughout the pores, because smaller molecules can enter the pores, they have a larger volume of liquid accessible to them. Molecules larger than the pores only have the liquid surrounding the beads accessible to them, and thus move through the column faster.

The smaller the molecular weight of proteins the slower they move through the ...

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