IBBR maintains a state-of-the-art NMR facility that includes NMR instruments with super-conducting magnets operating at 500 MHz, 600 MHz and 900 MHz proton resonance frequencies.
Understanding signaling across cellular membranes.
Integral membrane Proteins (IMPs) tightly control material transport into and out of cells, as well as initiate cellular signaling cascades in response to hormones, neurotransmitters and other environmental factors. These proteins are the material and informational gateway to the cell. IMPs are the target for ~50% of current FDA-approved drugs, emphasizing their critical importance to the pharmaceutical industry.
Biopharmaceuticals: Furthering development, manufacture and regulation of protein drugs.
In contrast to small molecule therapeutics, protein biologics are many kDa in size and require higher-order folding of the primary sequence (i.e., tertiary structure) for therapeutic function. While correct folding of a protein biologic is critical for drug efficacy, misfolding may impact drug safety by eliciting unwanted immune and/or other off-target responses. High resolution NMR provides a robust spectroscopic approach for obtaining structural “fingerprints” of the bioactive form(s) of protein therapeutics at atomic resolution. Such fingerprints of the structure(s) of protein therapeutics can provide an important tool for establishing consistency in drug manufacturing and for comparing a biosimilar to an innovator reference product.
The W.M. Keck/NIST X-ray Crystallography Core Facility provides consultation, collaboration and training in macromolecular structure determination. The Facility supports IBBR’s strong research emphasis providing experience and expertise in macromolecular structural determination by X-ray crystallography and analysis of the resulting atomic structures. The Facility’s staff can work with researchers who have an interest in incorporating macromolecular structure determination into their research projects through a variety of working relationships.
A crystallographic project has many components: initial analysis of the sample for purity and homogeneity, scanning of many possible crystallization conditions, refinement of the crystallization conditions to produce crystals suitable for structural analysis, preliminary exposure of the crystals to X-rays to determine whether they diffract well enough for structure determination, data collection, data processing, structure determination, structure analysis, and deposition of the structure into the Protein Data Bank. Researchers are encouraged to contact the Facility to discuss whether structural information could provide additional insight in their particular project.
The facility also provides training in crystallographic techniques.
- Two Rigaku MicroMax007 X-ray generators and two RAXIS IV++ detector equipped for cryogenic data collection.
- Phast gel electrophoresis, miniDawn laser light scattering, and MALDI mass spectrometry for analysis of protein samples prior to crystallization.
- Hydra II robot to set up crystallization screens.
- Personnel with many years of experience in growing crystals of biological macromolecules, collecting X-ray data, and determining 3-dimensional atomic structures.
- Access to crystallographic computer programs and multiple computer workstations
Small-angle x-ray scattering (SAXS)
The IBBR houses a SAXS instrument that allows measurement of biological molecules, complexes formed by these molecules and heterogeneous mixtures of these molecules in both the solution and solid state. This includes both dilute and concentrated solutions of proteins, nucleic acids, lipid nanodiscs, therapeutic viruses, and antibodies.