Biochemical Methods

In order to analyze an interaction of proteins of interest, the first thing to do is to produce and purify it. Using the proper expression vectors the respective protein will be synthesized in a bacterial culture and purified in two or three steps with different chromatographic methods. Standard techniques of molecular biology (in particular PCR, polymerase chain reaction) allow to construct the desired types of protein. First of all insertion of particular point mutation or deletion of one or more protein domains, followed by biophysical analysis of a modified properties and comparison with a wildtype permits to a significant interference on the functionality of the protein.

Protein purification

Cell harvesting

Centrifuges of different sizes allow the harvest of microbiological culture and separation of insoluble components after cell lysis.

Äkta Purifier

Chromatography anlage with different type of column for purification of recombinant protein from Bacterial lysat. The picture shows the Akta Purifier-System

UV/Vis- Spektroscopy

The essential requirements of a quantitative analysis of the protein interaction is exact definition of the concentration of involved interaction member. Using the UV/Vis- Spektroscopy (picture) we can utilize different methods (Gill&von Hippel, Bradford etc.)