New Strategies of Directed Evolution

Pluecki6

Many of the studies in the various projects required technologies that were simply not available at the time. Therefore, we have always been developing novel technologies to be able to greatly expand our toolbox, in order to make certain developments in molecular engineering possible in the first place.

One example is ribosome display, a method of cell-free selection and evolution from very large libraries. This approach has been adapted to affinity maturation (the current record is a monovalent dissociation constant of 1 pM), protein stability, or enzymatic turnover. Other challenging properties, such as the selection of new folds or robust topologies, are currently being investigated.

Another example is phage display, which has recently been adapted to make it compatible with a wider spectrum of proteins to be displayed (by changing the export pathway), and has also been used to select for properties other than mere binding, such as protein stability and enzymatic turnover.

A third example is the protein fragment complementation assay (PCA), which allows the selection of interactions inside the cell, in a very rapid manner.

Finally, technologies are currently being developed that deal with the selection of stable membrane proteins.