Protein Production & Biophysics (PPB)
The goal of the Protein Production and Biophysics (PPB) section is to produce purified proteins for investigators, for use other CBT sections (i.e. Structural Biology (SB) for NMR and crystallization and Target Validation & Screening (TVS) for use in in vitro assays), and for internal studies by the PPB section. The PPB has assembled the expertise to provide a wide range of methods for studying the biophysical properties of proteins including protein-ligand and protein-protein interactions.
The PPB program occupies modern laboratory space in the CBT within the Institute for Biosciences and Biotechnology Research (IBBR), a three-building complex of research laboratories at the University of Maryland College Park, Shady Grove in Rockville, MD and additional space at the University of Maryland School of Medicine in Baltimore, MD. It shares the facilities, equipment, and expertise with the other six programs in the Center for Biomolecular Therapeutics (CBT). In addition, the CBT has access to the core facilities available at the University of Maryland, Baltimore (UMB), the core facilities in the Center for Vascular and inflammatory Disease (CVID) at UMB, the shared services in the University of Maryland Marlene and Stewart Greenebaum Cancer Center (UMGCC), and the biophysics core facilities within the IBBR. All laboratories have refrigerators, freezers, house vacuum, hood ventilation, and natural gas hook-ups. Autoclave, dishwashing, and pure water facilities are located adjacent to the laboratory space. Office suites (PI, postdocs, students and staff) are clustered together and located adjacent to the laboratory space. All offices are equipped and with desktop computers and broadband internet access is freely available throughout the site (wired and wireless). The CBT/IBBR complex has a business office to assist with purchasing/receiving as well as on-site information technology support and facilities support.
The facility is capable of cloning and expressing tagged and untagged proteins in a wide range of scales from flasks to bioreactors in bacteria, yeast, baculovirus, or mammalian systems. In addition, uniform stable isotope (2H, 15N, 13C) and selective methyl labeling of proteins is regularly produced for NMR studies. The group is skilled in protein purification methods needed for tagged and untagged proteins including HPLC/FPLC techniques. The PPB group regularly performs molecular biology techniques (RNA/DNA purification, RT-PCR, cloning, stable and transient protein expression in cells. The PPB group has dedicated biosafety cabinets, CO2 incubators, inverted phase microscope, low/high speed centrifuges, and storage capabilities including liquid nitrogen storage freezers.
The program has expertise in the characterization of the proteins including stability, structure/dynamic properties, size, covalent modification (i.e. phosphorylation, ubiquitination, methylation, glycosylation etc). Services for the identification of unknown proteins are available using a multitude of biophysical/biochemical techniques including denaturing polyacrylamide gel electrophoresis (SDS-PAGE), FPLC, mass spectrometry (MS), isothermal titration calorimetry (ITC), nuclear magnetic resonance (NMR), differential scanning calorimetry (DSC), circular dichroism (CD), fluorescence, UV/Vis, DLS, analytical ultracentrifugation (AUC), kinetic stop-flow/quench-flow, and surface plasmon resonance (SPR) techniques. Expertise & equipment for measuring enzyme kinetics, binding kinetics, and thermodynamic properties of a enzyme/proteins are also available including monitoring binding to substrates, other proteins, ligands, and/or oligomerization properties.
David Weber, PhD
108 N. Greene Street, Room 439
Baltimore, MD 21201