Team Hart

Responsible : Darren Hart

The Hart team uses combinatorial molecular biology methods to study proteins and their complexes that are of biological and medical interest. Combinatorial methods (e.g. directed evolution, phage display) address problems that are too complex for rational design approaches. Mimicking evolution in a test tube, large random libraries of variants are screened to identify rare hits with the desired property.

Using structure-guided phage display we design and construct large libraries of potential interactors (peptides or protein domains) that are panned against the target to identify high affinity binders with applications as inhibitors. Current projects build on a long term interest in influenza virus polymerase to target viral replication via both virus and host targets.

In our ESPRIT process, for example, all truncations of a target protein are generated and screened using advanced picking and arraying robotics. Consequently, we are able to study certain biological questions with advantages over classical approaches. Historically, this opened up X-ray structural studies on the influenza polymerase and has now been applied to viral, human and plant proteins. Funded access to this unique technology is possible via Instruct-ERIC.

Members

Darren Hart (Research Director CNRS) (ORCID profile)
Philippe Mas (Engineer CNRS)
Alberto Florez-Prada (Ph.D. student)
Benoît Sépari (Ph.D. student)

Figure 1 : Screening tens of thousands of expression constructs of a target gene. Constructs are made as a random library and printed on membranes for soluble expression analysis by hybridisation of fluorescent antibodies.