Structural and functional analysis of the interaction between ICAM-1 and DBL domains implicated in cerebral malaria

Objectives

The main objective of this project is to define the molecular basis of the DBLβ-c2–ICAM-1 interaction and develop monoclonal antibodies capable of blocking this adhesion to inform therapeutic strategies.

As specific objectives, there are:

1. Raise monoclonal antibodies against DBLβ-c2 domains, test the ability of these antibodies to bind ICAM-1, and use them to select ICAM-1 binding parasite lines.
2. Test the ability of antibodies to inhibit the DBLβ-c2:ICAM-1 interaction and use mutagenesis and structural studies to characterise inhibitory antibodies.
3. Use mutagenesis to map the ICAM-1 binding site on the DBLβ-c2 domains.
4. Assemble complexes between DBLβ-c2 and ICAM-1 and use biophysical methods to study their stoichiometry and architecture.
5. Express and purify multiple ICAM-1 binding DBLβ-c2 domains and attempt to crystallise these domains alone, and in complex with ICAM-1 domains and Fab fragments from monoclonal antibodies.