Laboratory for the Study of Intermolecular Interactions

Protein Expression and Purification Laboratory
This lab provides high-purity protein samples suitable for structural studies. It uses both traditional and restriction-free cloning techniques to build expression vectors, leveraging a wide variety of bacterial plasmids to optimize protein solubility. For insoluble proteins, refolding protocols can be routinely optimized. The lab specializes in small-scale parallel screening of bacterial strains to improve protein quality and can scale up to 10L cultures to produce milligrams of recombinant proteins. Equipment includes thermostatic and refrigerated incubators, high-capacity refrigerated centrifuges, a high-speed centrifuge, and a sonicator for producing homogeneous lysates. Protein purification is performed using multiple chromatographic techniques (affinity, ion exchange, hydrophobic interaction, size exclusion) on an AKTA PURE 25 FPLC system. A dedicated cold room is available for temperature-sensitive targets. Under PRP@CERIC a UPLC-MS instruments, Waters BioAccord, has been acquired and introduced into the protein production pipeline, as a powerful instruments for protein characterization.
Biophysical Characterization Laboratory
This facility hosts instruments for analyzing the biophysical properties of proteins and intermolecular interactions. Techniques include:
• UV-Visible spectrophotometry
• Dynamic Light Scattering (DLS) for assessing molecular size and polydispersity
• Differential Scanning Fluorimetry (DSF) for thermal stability and high-throughput protein-ligand interaction screening
• Isothermal Titration Calorimetry (ITC) using the MicroCal iTC200 and PEAQ-ITC for label-free, in-solution binding analysis, delivering full thermodynamic profiles (KD, stoichiometry, ΔH, ΔS) under native conditions
• Grating Coupled Interferometry (GCI) using the WAVE delta, a fast, label-free biosensor using microfluidics and waveRAPID to analyze kinetics and affinity across a wide range of molecules, including large complexes. It handles crude samples, delivers high-resolution data quickly, and is ideal for drug discovery and challenging interaction studies.
• Switchsense DNA nanolevers using the heliX+, a system that employs fluorescent DNA nanolevers to study biomolecular interactions in real time. It measures binding kinetics, conformational changes, and enzyme activity with high sensitivity. Multiplexing and two-color detection allow simultaneous analysis of multiple interactions or binding modes.
• Absorbance based assays (ELISA, colorimetric assays); Fuorescence based assaya (Fluorescence Anisotropy, Time-Resolved Fluorescence, HTRF®, LanthaScreen®, 3D spectral scans); luminescence (Glow, flash, multicolor assays, spectral scans, BRET™, DLR™); AlphaScreen based (AlphaScreen®, AlphaLisa®, AlphaPLex®) by the Spark® Microplate Reader, a versatile, high-throughput plate reader that supports low-volume samples and full automation for consistent, multipurpose detection across various assay types.
XRD1 Beamline at Elettra Synchrotron
XRD1 is a multipurpose diffraction beamline for macromolecular and small molecule crystallography. It features a multipole wiggler source (4–21 keV range) and a double-crystal monochromator optimized for anomalous signal detection. It supports single-crystal diffraction at cryogenic temperatures, powder diffraction with temperature control, and surface diffraction in grazing incidence using laser-assisted alignment and helium path.
Instrumentation includes:
• Huber kappa goniometer with a Dectris Pilatus 2M hybrid 2D detector (90–600 mm sample-detector distance)
• Oxford Cryostream 700 cryostat (temperature range: 80–400 K)
• Automated sample changer and data collection robot