NovAliX Workshop | Monday, May 9 | 16:15
Combining biophysics to enable drug discovery: showcasing structural MS and SPR
Presenters: François Debaene & Renaud Morales
NovAliX is a drug discovery-focused CRO with a wide range of services that support projects from hit discovery to optimized leads. With around 250 researchers, the drug discovery team has extensive expertise in computational and medicinal chemistry to drive discovery programs. This capability can be combined with different screening platforms such as FBDD, covalent fragment screening and DEL for hits and their development. NovAliX has a unique biophysical platform operated by passionate expert scientists covering MS, SPR, NMR, MST, DSF. Structural information can be provided by protein X-ray crystallography and cryo-EM - including membrane proteins. NovAliX offers strong capabilities in synthetic chemistry and chemical process research.
In this workshop, we will see through illustrative examples the use of various biophysical techniques and how they have been instrumental towards an advancement in biological understanding of the nuclear receptor target ROR''.chr('61543').'', hence accelerating its drug development program. Particularly, Structural MS (Native MS, HDX-MS) and SPR, two techniques very well mastered by our teams, will be used as a source of information in the illustrated examples.
Dynamic Biosensors Workshop | Tuesday, May 10 | 16:05
switchSENSE® heliX®: measuring interactions from small molecules to cells
Presenters: Daisy Paiva & Alice Soldà
switchSENSE® is an automated, fluorescence-based biosensor chip technology that employs electrically actuated DNA nanolevers for the real-time measurement of binding kinetics (ka, kd) and affinities (with KD values down to the fM range). The platform offers an automated ligand density control, which allows to conveniently discriminate between affinity and avidity in one single assay. Interactions between proteins, DNA/RNA, and small molecules can be detected with femto-molar sensitivity. At the same time, conformational changes and complex binding events can be measured using minimal amounts of sample. Using heliX® biosensor and the novel DNA Y-structure, we can characterize ternary complex formation of bifunctional small molecules, like PROTACs and molecular glues. The (homo- or hetero-) target proteins can be functionalized on the end of the two FRET pair color-coded Y-arms of the structure. The Y-structure closes upon small molecule binding, and the subsequent ternary complex formation brings together the green donor and the red acceptor dye into a closer, FRET-sensitive, distance. Herein, we demonstrate how we can characterize kinetic rates and compound ranking in a high-throughput manner, gaining information on binary and ternary complex formation in one experiment. The sensitive FRET read-out can be also used to analyze in real time other complex binding modes, like the kinetics of 2-domain RNA-binding proteins (RBP) with two different binding motifs within the ssRNA overhang placed on the DNA biochip.
Moving from small molecules interactions to complex systems, we present a novel method to capture isolated cells and to measure the association and dissociation kinetics of fluorescently labeled antibodies to/from cell surface antigens in real-time. To this end, flow-permeable, mesh-like cell cages were designed to accommodate and physically retain single in-suspension or adherent cells in the microfluidic channel of a commercially available biochip. Herein, we present the first kinetic analysis of bispecific T cell engagers (BiTE®) binding to T cell and cancer cells.
In this work, we highlight the recent advances that allows investigation of biomolecular interactions under conditions that are close to in-vivo situations in a highly automated workflow, and thus useful for the screening and characterization of new drugs that intervene in cellular or biochemical processes.