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Biotechnology Bioprocess Engineering Measurements Biomolecular Materials Research |
Division Contact: Vincent Vilker Working in several areas of DNA technologies, we are developing methods for measuring DNA damage and repair, for characterizing DNA including mutation detection, genetic toxicology, human identity profiling, linearly arrayed DNA on solid substrates, and developing human DNA Standard Reference Materials®. We are developing experimental methods and standards to measure DNA damage and its repair in mammalian cells exposed to free radicals generated by ionizing radiation, carcinogenic compounds, or redox-cycling drugs. Free radicals produced in vivo are thought to be mutagenic and carcinogenic, and are associated with numerous diseases such as cancer. Measurement of DNA damage at the molecular level in mammalian cells is a prerequisite to understanding the chemical mechanisms of damage by free radicals. We also are characterizing enzymes involved in DNA repair. DNA repair enzymes are essential for preserving the integrity of the genetic material and are potential anti-viral and anti-cancer drug targets. Techniques used include gas chromatography/mass spectrometry (GC/MS) and liquid chromatography/mass spectrometry (LC/MS). We are working on new methods for DNA profiling, ranging from developing well-characterized DNA standards for restriction fragment length polymorphisms to performing research for rapid determination of DNA profiles by polymerase chain reaction amplification and automated detection of fragments. In addition, we are interested in cooperative development of short-tandem repeat and single nucleotide polymorphism detection technology, multiplex polymerase chain reaction, DNA stability studies, Y chromosome markers, mitochondrial DNA sequencing, and attendant standards. Techniques include sensitive staining of electrophoretic gels, use of chemiluminescence, enhanced applications of capillary electrophoresis, and matrix-assisted laser desorption ionization time-of-flight mass spectrometry. We are developing standards and methodologies for mutation detection, a burgeoning area in clinical laboratories. At present, national standards are not available for heritable or cancer-related genetic testing. We are focusing on standard materials intended for DNA-based mutation detection systems for diagnostic testing and to analyze research parameters that affect measurement quality in various assay systems. We are validating novel DNA-based biomarkers for early detection of cancer in collaboration with a consortium of universities and other governmental agencies. We are developing biomarkers, measurement technologies and standards for tissue-engineered materials. These will help assure that tissue-engineered materials are free of biological and genetic changes capable of compromising their safety and viability. We are interested in the cooperative development of oxidative DNA damage and chromosome damage markers, cellular inflammation detection, microarrays, tissue-engineering biomarkers, and attendant standards. Techniques used include GC/MS, LC/MS, fluorescence in-situ hybridization, gene expression and real-time diagnostic polymerase chain reaction. Contact: Miral Dizdar Detailed knowledge of the structures and dynamics of macromolecules is critical for a complete understanding of biochemical reaction mechanisms and molecular recognition processes. Such an understanding can impact bioengineering efforts in the areas of biomaterials, "designer" enzymes, and pharmaceuticals. We are developing measurement methods, models, and databases needed to study macromolecular structure and dynamics using nuclear magnetic resonance (NMR) spectroscopy. NIST and the Center for Advanced Research in Biotechnology maintain a state-of-the-art NMR facility, which includes superconducting magnets operating at 500 megahertz and 600 megahertz proton resonance frequency. Research projects include investigations into the structure and dynamics of redox proteins, DNA enzyme-ligand complexes, RNA and DNA oligonucleotides, nucleic acid binding proteins, nucleic acid-protein complexes, and signal transduction proteins. We also are using NMR measurements to study protein folding. Contact: John P. Marino Bioprocess Engineering Measurements We are focused on the development of measurement methods, databases, and generic technologies related to the use of biomolecules and biomaterials in manufacturing. We have developed measurement methods and data in the protein biospectroscopy area that will lead to improved understanding of intra- and interprotein electron transfer processes. This understanding helps industrial biocatalyst development by allowing for more efficient utilization of carbon sources (e.g., renewable resources) and nutrients, and in developing new ways to drive organic syntheses such as the stereospecific hydroxylation of pharmaceutical precursors. We also are using biospectroscopy measurements to develop Standard Reference Materials® for use in calibrating the intensity of fluorescence signals in many fields of biology. In the biothermodynamics of enzyme-catalyzed reaction project, we are combining chromatography and microcalorimetry measurements with chemical equilibrium analysis to develop thermodynamic data for several industrially important biotransformations. Currently, we are focusing on the metabolic pathway by which micro-organisms and plants convert glucose to aromatic amino acids. Several large chemical companies are investigating this chorismate metabolic pathway as an environmentally friendly source of aromatic hydrocarbons. A new collaborative project among NIST, the U.S. Department of Agriculture, and the European Commission Institute for Reference Materials and Measurements will focus on developing reference materials for use in the detection of genetically modified crops. In another project, we are applying electrochromatographic/electrophoretic separation equipment and methodology to the separation of different physical forms of DNA (supercoiled plasmid, relaxed circular plasmid, and linear genomic). Contact: Vincent L. Vilker Biomolecular Materials Research A current trend in materials development is to employ biological molecules, biological principles, or both. Such materials are sometimes referred to as "biomimetic," indicating they have characteristics such as self-assembly, molecular recognition, specific chemical responses, and complex molecular architecture, which lead to unique structural or functional characteristics. Chemically controlled biomimetic surfaces are essential components of biosensors, bioelectronics, biocatalytic systems, and many diagnostic devices. Biomolecular materials thus influence diverse applications such as health care, environmental pollution monitoring, agriculture, and chemical manufacturing. An underlying need for these applications of biotechnology is the characterization and control of biomolecules at interfaces. Fundamental studies are being conducted to better understand the structure and function of natural and biomimetic materials that self-assemble. Lipid membranes are one such class of self-assembling materials. They organize and control the structure of proteins that naturally reside within them, many of which have commercially important functions. We also study models of cell membranes as tools for achieving better quantification of therapeutic agents, which are likely to act at the level of the cell membrane via cell surface receptors or which have to pass through the cell membrane to be effective. Interactions among biological molecules and between biological molecules and surfaces occur during sensor operation, diagnostic tests, cellular recognition events and mobility, and in the formation of modified surfaces as organized biomolecular materials. In some of these applications, it is desirable to enhance strong molecule-specific interactions while minimizing weaker non-specific interactions. In other cases, many simultaneous weak interactions are needed to effect the appropriate dynamic response. We develop both the experimental tools, such as chemically controlled surfaces and techniques for monitoring reactions at surfaces, and the theoretical tools to improve understanding of dynamic biomolecular processes and permit predictions and optimization of reactions of biomolecules. We also are developing methods for kinetic analysis, current noise analysis, and stochastic models of processes at surfaces. We are examining
macromolecules including bacteriorhodopsin, DNA, and enzymes in various
configurations as potential components of electrochemical, electronic,
and optical devices. We engineer prototype devices to allow consideration
of real world issues such as fabrication and adaptation of instrumentation
and methodology to field conditions. Contact: John
Kasianowicz Center for Advanced Research in Biotechnology The Center for Advanced Research in Biotechnology (CARB) in Rockville, Md., was established by NIST, the University of Maryland, and Montgomery County, Md., to study structure and function relationships of biological macromolecules. CARB researchers are focusing on the measurement of structure by X-ray crystallography and nuclear magnetic resonance spectroscopy (NMR) as well as the manipulation of structure and activity by a variety of modern molecular biological techniques. Scientists use modeling,
molecular dynamics, and computational chemistry to understand protein
structure and are developing theory to predict the effects of specific
structural modifications on the properties of proteins and enzymes. A
variety of physical chemistry methods are used to measure and analyze
structural changes, activities, and thermodynamic behavior of biological
macromolecules. CARB maintains state-of-the-art facilities for X-ray crystallography,
NMR spectroscopy, molecular biology, and physical biochemistry. Contact: Edward Eisenstein
Date
created:
September 28, 2001 |