Dhaval Sangani

Remit was to establish diverse tools and platforms for TCR functional avidity evaluation and/or antigen discovery for ‘off-the shelf’ cell therapy applications.

Led development of a novel Mammalian Display platform for discovery &/or affinity maturation for high-throughput screening and engineering of functional and specific TCR candidates for bispecific therapeutics.

TCR bispecific discovery and development using phage display and protein engineering techniques for immunotherapeutic drug development.

Protein Charaterisation/Purification Mechanism of Action Studies Designing novel purification paradigms and Identification of protein binding targets of small molecules by employing suitable protein enrichment paradigms in conjunction with affinity-guided activity assays to correlate functional activity with protein abundance by MS. Provide biochemical support to quantitative proteomics MS platforms.

Publications • ‘Phosphotyrosine-dependent in vitro reconstitution of recombinant LAT-nucleated multiprotein signalling complexes on liposomes’ Sangani D, Harder T, Mol Membr Biol. 2008 Dec 29:1-13. • ‘Plasma membrane rafts engaged in T cell signalling: new developments in an old concept’ Harder T, Sangani D, Cell Commun Signal. 2009 Sep 4; 7:21. • ‘HIV entry in macrophages is dependent on intact lipid rafts’, Carter G, Bernstone L, Sangani D, Bee J, Harder T, James W. Virology. 2009 Jan 29. .

Engineering and expression of integral/membrane anchored and tyrosine phosphorylated Linker for activation of T cell (LAT) for in-vitro functional reconstitution studies. • Detergent extraction of tyrosine phosphorylated LAT from crude homogenates and enrichment by immunoaffinity beads. • Characterisation of T cell membrane protein-protein interactions by affinity analyses (Surface Plasmon Resonance) of recombinant SH2, SH3 domain and immunoglobulin fold proteins (expressed in bacterial/CHO/ insect cells) • Recombinant protein production for X-ray crystallisation and production of N15-labelled protein for NMR. • Membrane protein expression and engineering for reconstitution of multiprotein complexes on artificial liposomes. • Stable transfection of mammalian cell lines with live colour vector constructs for fluorescence imaging studies on membrane signalling machinery.