The Protein Man's Blog | A Discussion of Protein Research

Affinity chromatography is the most popular and widely used method for purification of biomolecules including proteins, antibodies, lectins, carbohydrates, nucleic acids and glycoproteins. Affinity purification is based upon specific surface interactions among biological molecules such as antigen-antibody, enzyme-ligand etc. These specific surface interactions enable the purification of molecules from 1000 to 10,000 fold in a single step. In affinity chromatography, the ligand, antigen or one of the interacting molecules is covalently bound to the matrix and is used as a bait to fish out the target protein from any complex pool of proteins including cell or tissue lysate.

Extracting proteins from plant and animal sources are two different things. While animal proteins can easily be extracted, working with plant proteins can be quite a challenge. Why? Plant proteins are more difficult to extract since they are protected by a rigid cell wall that contains several interfering agents (i.e proteases).

As mentioned in our earlier posts, extracting, separating and identifying proteins from plant samples present a unique challenge since they contain significant amounts of proteases and other compounds that may interfere with the process. Generally, there are three critical aspects for successful protein separation:

Cell disruption is a collection of techniques used for releasing biomolecules of interest from inside the cell.  This blog will discuss the various techniques which will be broken down into two categories, gentle and harsh cell disruption, there relative advantages and which tissue types work best with the particular technique.