ThermoFisher Scientific/CCG Pilot Project awards announced
January 10, 2007
ThermoFisher Scientific in conjunction with the University of Michigan Life Sciences Institute is pleased to announce the awards for its second annual Center for Chemical Genomics Pilot Project Initiative. The funds will provide research support for up to two years, allowing University of Michigan scientists to conduct innovative research directed toward technology development relating to high throughput screening, small molecules synthesis, target detection and assay miniaturization.
Researchers receiving awards are Assistant Professor of Pathology Jason Gestwicki for "Systematic Generation of Uniformly Modified Drug Libraries."
Professor of Electrical Engineering Jay Guo for "Label-free Optical Micro-Resonator Biosensor for High-throughput Screening Applications."
Assistant Professor of Materials Science and Engineering Jinsang Kim for "Polydiacetylene-based Self-signal Amplifying Protein Sensors and Microarrays."
Assistant Professor of Biomedical Engineering and Chemical Engineering Michael Mayer for "Efficient and Parallel Formation of Membrane Protein Arrays by Hydrogel-based Microcontact Printing."
Professor of Chemistry John Montgomery for "New Methods for Macrocycle Glycosylation."
Abstracts
Jason Gestwicki, Ph.D.
Assistant Professor, Department of Pathology
Research assistant Professor, Life Sciences Institute
Systematic Generation of Uniformly Modified Drug Libraries
Protein-protein interactions have proven difficult to block using traditional chemical approaches but they are potential targets of numerous diseases, such as neurodegenerative disorders and cancer. The primary goal of this project is to develop synthetic methods that enhance the ability of small molecules to inhibit protein-protein interactions. Briefly, this approach involves coupling of a small molecule to a chemical functionality that has high affinity and selectivity for FK506-Binding Protein (FKBP). After coupling, the molecule gains the steric bulk of FKBP and this drug-protein complex is designed to be a more substantial barrier to additional protein contacts. Indeed, we have recently shown that bifunctional derivatives of an amyloid ligand are 40-fold more potent when they are attached to FKBP-binding groups. Our approach is inspired by the natural binding mode of rapamycin and FK506 and it relies on the use of a native, endogenous protein to create semi-synthetic inhibitory complexes. In this proposal, we will develop the high yielding and selective chemistry required to expand the scope of this concept to large-scale parallel syntheses. These reagents and concepts are designed to expand the utility of the CCG’s existing chemical library and enrich the resulting compounds for inhibitors of protein-protein interactions.
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L. Jay Guo
Professor of Electrical Engineering
Label-free Optical Micro-Resonator Biosensor for High-throughput Screening Applications
We propose to continue developing novel optical system for studying real-time protein binding interactions and for high-throughput screening of chemical compounds that are important in protein-protein interactions. The method is based on optical microresonators (OMR) of very high quality-factor acting as highly sensitive reporter of biomolecular binding events. These microresonators, designed with integrated optics techniques, are formed using closed-loop (ring type) shaped waveguides. A typical dimension of such a microresonator is 20µm to 100μm depending on the optical wavelength and other design parameters. Our previous measurements showed a high sensitivity for detecting glucose concentration and biotin-streptavidin binding. In year 2 we plan to carry out many screening experiments using relevant protein systems important to the researchers at CCG. In addition, the year 1 program also allowed us to exploit a new sensor platform that uses commercially available glass microtube as a new OMR sensor platform. This new approach eliminates the device fabrication process and our initial results showed very high resonance quality factors. More importantly, this new platform naturally incorporates the fluid handling capability in itself without additional microfluidic elements. We will investigate its full potential in the year 2 program. We plan to pursue these two platforms simultaneously since each has its distinctive advantages. The grant will help to establish that OMR sensor arrays have the potential of providing high-throughput analysis of biomolecular interactions without fluorescently tagging the analyte molecules.
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Jinsang Kim, Ph.D.
Assistant Professor
Materials Science and Engineering, Chemical Engineering
Macromolecular Science and Engineering, and Biomedical Engineering
Self-signaling and Signal Amplifying Conjugated Polymer-based Biosensors and Sensor Arrays
Fast and reliable detection of diagnostically important biological molecules remains a significant challenge especially given the difficulty in devising an effective label-free and sensitive detection strategy. Rationally designed conjugated polymers have provided high sensitivity by responding sensitively to a variety of environmental stimuli via changes to their physical properties. In this project we will combine the signal amplifying property of synthetic conjugated polymers with the unique specificity of biological molecules in recognition. To achieve this it is desirable to devise bio-conjugated polymer hybrid materials where biological materials are the recognition receptor and CPs are the reporters generating a sensory signal. The most important design principle to address is the way to combine a biological receptor and a conjugated polymer so that a selective recognition event at the receptor site can produce a sensitive, amplified signal from the conjugated polymer reporter. Based on our preliminary results on self-signal amplifying DNA sensors and sensor arrays this pilot proposal seeks to advance the understanding of the molecular design principles of conjugated polydiacetylenes in order to develop self-signaling and signal amplifying protein microarrays for sensitive protein detection and drug development and screening. The anticipated results from the proposed research will allow a universal biosensor design principle to realize amplified label-free detection of biological molecules and drug screening. This will eventually contribute to promoting public health by providing efficient early diagnostic screening of disease and facilitating drug development and screening.
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Michael Mayer
Assistant Professor of Biomedical Engineering and Chemical Engineering
Efficient and Parallel Formation of Membrane Protein Arrays by Hydrogel-based Microcontact Printing
In continuity with our previously funded grant proposal, we propose to develop an efficient technique to fabricate arrays of membrane spots containing various membrane proteins using hydrogel-based microcontact printing. The emphasis of this follow-up proposal lies in expanding the number of membrane proteins, and, most importantly, in rendering the technique practical for wide-spread use.
The advantage of the hydrogel stamping method is that hydrogel posts on the stamps can store fragile biomolecules (in this case, lipids and membrane proteins) in a hydrated and biocompatible environment and that they can deliver those molecules to substrates at well-defined locations. These unique capabilities of hydrogel stamps make it possible to create rapidly many arrays of membranes while consuming extremely small amounts of precious molecules such as transmembrane proteins.
During the past nine months of funding, we developed a technique to employ hydrogel stamps for printing arrays of membrane proteins. Due to the low conc. (as well as the extremely high cost) of proteins from commercially available sources, however, fabrication of multiple copies of the same array is challenging.
Here we propose to continue this work towards implementation of a practical, automated, and cost effective system for fabrication of membrane protein microarrays for high-throughput screening (HTS) applications. To miniaturize and automate the system, we propose to use a microarrayer to ink hydrogel stamps with small post size (≤ 100 μm). Moreover, to reduce the costs, we propose to grow commercially-available cell lines that over-express membrane proteins of pharmaceutical interest such as G protein coupled receptors (GPCRs) in our lab and then to employ nitrogen cavitation to obtain membrane vesicles with high protein concentrations in a cost-effective and readily expandable fashion (by growing additional cell lines). We will examine the functionality of these arrays using ligand-binding assays. We expect these studies to be instrumental in accelerating drug discovery.
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John Montgomery
Professor of Chemistry
New Methods for Macrocycle Glycosylation
The principal aim of this proposal is to devise a new procedure for the glycosylation of the complex molecules that may be constructed by the nickel-catalyzed multicomponent coupling procedures discovered in our laboratory. The exquisitely selective molecular recognition imparted by carbohydrate units coupled with the tremendous structural diversity in complex molecules that our nickel cyclization technology allows provides significant opportunities for the preparation of glycoconjugates with a diverse array of biological properties. In order to take full advantage of the potential chemical diversity that may be obtained by the merger of our cyclization technology with glycosylation methods, we envision that a two step method for assembly of a complex macrocyclic structure and stereoselective glycosylation, potentially without protecting groups, could have enormous impact in many avenues of research in chemical biology. Extension to classes of compounds including synthetic small molecules and peptides could have far-reaching consequences. In order to pursue this ambitious objective, we propose the following specific aims:
- To continue to refine our newly discovered procedures for the synthesis of glycosylated macrocycles from ynals or ketones.
- To develop strategies for the site-selective glycosylation of complex natural products.
- To devise strategies for the synthesis of libraries of sugar conjugates of synthetic small molecules and small peptides and to demonstrate the approach with representative sugar silane reducing agents.
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