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For information, please contact Mario Bianchet email@example.com
We have a an array of instruments that are available for use by members of the Johns Hopkins community as well as by outside users. Equipment includes:
Surface Plasmon Resonance (SPR) interaction analysis equipment (Biacore 8K) will provide a label-free detection method for studying the binding behavior of biomolecules. The Cytiva™ Biacore 8K is a high-throughput and high-sensitivity SPR system that delivers binding data of outstanding quality. Samples are immobilized on a chip surface using one of several available chemistries while both binding and dissociation of another molecule are monitored. This method can be used to study a wide array of binding interactions.
Mass photometry (MP) is a revolutionary new label-free method for accurately measuring the mass of single macromolecules in solution. MP builds upon the principles of interference reflection microscopy and interferometric scattering microscopy. As a result, the light scattered by single molecules can be reliably detected and correlated to their molecular mass. The Refeyn® TwoMP™ mass photometer provided by the facility will allow the characterization of the biomolecular samples, their oligomerization, biomolecular interactions, and macromolecular assemblies.
MicroScale thermophoresis (MST) – NANOTEMPER® Monolith™ NT.115. This instrument allows a straightforward, fast, and precise approach to quantifying biomolecular interactions. MST is the exception to the label-free techniques instruments of the core. One of the molecules must be fluorescently tagged. Its advantage is that it allows the detection of a broader range of interaction types, ranging from ion and fragment binding up to interactions of macromolecular complexes such as liposomes or ribosomes. MST has become a standard technique to detect specific macromolecule-probe interactions. This method measures the differences in the movement rate through a microscopic temperature gradient caused when complexes are formed. MST can be applied to characterize any biomolecular interaction, including protein-DNA, protein-RNA, protein-protein, and antigen-antibody interactions, as well as ligand binding to ternary complexes.
Size exclusion Multi-Angle Light Scattering (SEC-MALS) Detector. SEC-MALS combines size-exclusion chromatography with multi-angle light scattering to determine molecular mass and size in solution, offering an advanced characterization technique and overcoming the limitations of size-exclusion chromatography-column calibration. A Wyatt DAWN® HELEOS™ Light Scattering detector Instrument is connected to a size exclusion column attached to an Agilent high-performance liquid chromatography (HPLC) instrument. Combining a UV/Vis absorbance detector with a HELEOS and Optilab refractive index detector creates a UV-MALS-RI triple-detection system to analyze copolymers and protein conjugates (glycoproteins, PEGylated proteins, surfactant-bound membrane proteins, etc.). With the addition of a WyattQELS DLS module for size determination, protein conformation may be analyzed. The DAWN may also be used in batch (off-line) mode to characterize the weight-average molar mass, z-average size, and second virial coefficient of unfractionated samples.
This instrument allows the detection of molecules in the range of 200 Da to 1GDa with molecular size (ratios of gyration, Rgs) between 10 to 500 nm and up to 1000 nm with shape-specific models and molecular sis range using combined Rh with DLS by flow 0.5 to 300 nm and by batch between 0.5 nm to 1µm with a sensitivity of 200 ng of BSA (in PBS)
Isothermal titration calorimetry (ITC) and. MacBio core will provide two different choices of calorimeters: an ITC VPITC and a Malvern Analytics ITC200, which handles smaller sample sizes (200 µl). ITC is the gold standard for quantitating molecules’ interactions and is used to study a wide range of biomolecular interactions. Applications range from drug design to fundamental research, such as understanding and regulating signal transduction pathways.
Differential Scanning Calorimetry (DSC) is an analytical technique for characterizing the thermal stability of biomolecules. It does this by measuring the heat change associated with the molecule’s thermal denaturation when heated at a constant rate. DSC is also used to determine the change in heat capacity. DSC can elucidate the factors contributing to the folding and stability of native biomolecules. The facility has a Malvern Microcal VP-CAPILLARY DSC Micro Calorimeter. These systems deliver direct, label-free, and in-solution measurements of binding affinity and thermodynamic parameters in a single experiment.
Tray-setting robot. The SPT Labtech Mosquito® is a liquid-handling robot that can set up 48- and 96-well crystallization trays using nanoliters of protein solution per drop. The instrument uses positive displacement pipetting using disposable micropipettes with individual pistons to deliver exceptional accuracy and precision, irrespective of liquid class. In addition, the instrument allows accurate, reproducible automation for liquid cubic phase (LCP) setups of 20 nl.
Automatic imaging of crystal trays. The Formulatrix Rockimager® automatically records images of crystallization drops while maintaining crystal trays at a constant temperature. High optical resolution is achieved by increasing the numerical aperture of the objective and combining multiple slices into one Extended Focus Image. The instrument also facilitates record-keeping for the barcoded crystal trays. Multiple images of each drop can be captured with user-adjustable settings, including exposure, polarization, and condenser aperture.