

Spring 2005 – SPECIAL ISSUE C3B Reporter An Annual Communiqué
In This Issue · C3B – Five Years in the Making · In the News · State of the Biosensors Lab · State of the Biochips Lab · State of the Signal Processing Lab Links Projects List of C3B projects and status Publications List of research papers submitted for publication Presentations List of presentations made by C3B researchers Other News Proposals, patents, and other news People Meet two of the C3B researchers, the Summer Interns, and the new Administrative Director Contact Us http://www.biochips.org fongr@vcu.edu A quarterly report to the faculty and industrial members on activities and developments within the C3B	C3B – Five Years in the Making The Center for Bioelectronics, Biosensors and Biochips was officially dedicated in October, 2000 as a center for research and development of engineered biosystems. In less than five years, C3B has established a well deserved reputation for innovative and multidisciplinary research in the VCU School of Engineering. C3B’s collaborative culture fosters a team-oriented research approach that has been exemplified in projects with the School of Medicine and the Department of Chemistry in the College of Humanities and Science. Exciting research is being conducted in the Biosensors and Biochip labs. This special issue of the C3B Reporter highlights the key projects, updates the news on the 4P’s (publications, presentations, proposals and patents), and introduces the researchers who are making it happen. Plans are in place to continuously improve C3B’s relationships with its constituencies, and to expand its base of support. At the leadership helm of the C3B is Dr. Anthony Guiseppi-Elie who was quoted in the Richmond Times-Dispatch, "I have the best job in the world." C3B In the News (Click on hyperlinks to access) C3B will be hosting its Annual Meeting on Thursday, June 2nd, 2005 at VCU School of Engineering. For more information or to register for this important event, please visit C3B Spring 2005 Meeting. VCU's 'global player' A professor in two departments, he founded biotechnology company and is widely known for research… RICHMOND TIMES-DISPATCH, November 2, 2004 “NanoBiotechnology: What’s All the Fuss about Small Stuff” was the subject of Professor Guiseppi-Elie’s keynote address at the Virginia Space Grant Consortium Scholars and Fellows Luncheon. The luncheon, hosted by Virginia Tech President Steger at the Newport News Omni Hotel on April 1, 2005, celebrated the accomplishments of the Consortium’s graduating Fellows and Scholars. Guiseppi-Elie, the man and his mission, to be profiled in the June 2005 issue of BioTechniques -- look out for this issue. State of the Biosensors Lab “We are moving into the animal testing phase of the implantable biosensor for glucose and lactate monitoring,” said Dr. Sean Brahim, Research Associate and Lab Director. In this high profile Department of Defense funded project, the next step is working with the School of Medicine to test the feasibility of the biosensors by implanting them into lab animals. The in vivo biosensor for the continuous monitoring of tissue glucose and lactate has important applications in risk stratification in trauma care, in chronic diabetes care and is being developed for military battlefield management of wounded soldiers. Other key projects in the Biosensors Lab include the development of an impedimetric biosensor for the monitoring of urea for dialysis management, the development of novel biomimetic and responsive (smart) polymeric hydrogels and the application of single walled carbon nanotubes for reagentless nanobiosensors. State of the Biochips Lab G. Scott Taylor, Ph.D. (engineering) student, Acting Lab Director, “We have a state-of-the-art laboratory that is well equipped to conduct our gene expression and biochip experiments.” (Note: see equipment acquisition) The Biochips Lab has a Class 1000 clean room for producing DNA microarrays, along with the instruments necessary for conducting gene expression and DNA hybridization experiments. Current projects with Pathology (Dr. Carleton Garrett) involve examining brain, ovarian and breast tumor samples; with Neurosurgery (Dr. William Broaddus) involve the study of the genetic anatomy of primary brain tumors, and with Chemistry (Dr. Nick Farrell), a study of platinum-based drug candidates for their physicochemical interactions with DNA. State of the Signal Processing Lab The Signal Processing Laboratory is a virtual laboratory vested with the responsibility for applying appropriate SP tools and techniques to all of the biosystems metrology tools under development at the C3B. “Signal processing and informatics are central to our mission, and, is one of the underpinning disciplines to our Center”, said Center Director, Anthony Guiseppi-Elie. We were saddened by the recent departure of Dr. Arvind Srivastava, the former director of this laboratory, who has moved to join the nanotechnology center, NUANCE, at Northwestern University, Evanston, Illinois. "We now have an opportunity to redefine this position in view of the broad and varied undertakings at the C3B" said Guiseppi-Elie. This laboratory will now be called the Signal Processing and Informatics Laboratory and will embrace our bioinformatics tools and platforms as well as our traditional signal processing capabilities. We will be looking to appoint a suitable postdoctoral candidate with breath of experience and knowledge in SP and bioinformatics.

Current Projects

Biosensors Lab

· Implantable Biosensors and biocompatibility of bio-smart materials

· Impedimetric Urea Biosensor

· Cell-based Neurotoxicity Array Biosensor

· Electronic NOSE for Trauma Monitoring

· Impedimetric and Amperometric Targeted Array Genosensor

Biochips Lab

· Class prediction of Astrocytomas

· Platinum Chemotherapeutic (PC) Inhibition of DNA degradation

· Microarray Cross-Platform Comparison

· Brain tumor heterogeneity of gene expression

· Preclinical Investigation of Mechanism of Action of a Novel Platinum Compound BBR3464

Completed Projects

· Impact of surface chemistry and blocking strategies on DNA microarrays

· A Factorial Study of the Influence of Production and Protocol Variables on Gene Expression Data From Spotted

Publications

Two important papers on achieving biological "stealth":

In two landmark papers, Prof. Guiseppi-Elie and his students describe the molecular engineering of biomimetic synthetic polymers that are capable of achieving biological stealth when implanted into the body of rats. That is, fooling the body to believe that an implanted device or material is indeed part of native tissue. In their recently publish paper that appeared in Biomaterials (Molecularly Engineered p(HEMA)-based hydrogels for implant biochip biocompatibility, Biomaterials (2005) 26: 4767-4778) graduate student Sheena Abraham, research associate Dr. Sean Brahim and Prof. Anthony Guiseppi-Elie, in collaboration with Prof. Kazuhiko Ishihara, The University of Tokyo, Japan, describe the details of design and synthesis of biomimetic materials based on the use of phosphorylcholine and polyethylene glycol. Phosphorylcholine (PC) is the active head group of the outer leaflet of the membrane bi-layer of most cells. The chemistry of this group was imparted to synthetic hydrogels, commonly used to make plastic contact lenses. In this seminal paper, the molecularly engineered hydrogels were demonstrated to resist common ECM protein adsorption while remaining non-cytotoxic to human epithelial and fibroblast cell cultures. These materials therefore exhibit the much desired and sought-after characteristics for implant material coatings. In another related manuscript submitted to the journal Langmuir, (Molecularly Engineered p(HEMA)-based Hydrogels: Hydration, Protein Adsorption and Cytotoxicity), the details and kinetics of protein adsorption are described and modeled in relation to the extent of hydration of these hydrogel materials. This research is supported by a grant (PR023081) from the US Department of Defense (DoDPRMRP).

Carbon nanotubes enable direct nano-biosensors for Enzymes and Proteins - NanoBiotechnology paper:

In 2001 Prof. Guiseppi-Elie and postdoctoral fellow, Chenghong Lei , along with University of Texas collaborator, Raymond Baughman published a paper on the use of single-walled carbon nanotubes as substrates for direct electron transfer to the flavin centers of oxidoreductase enzymes. This paper has achieved notable citations that place it in the top 1% of papers in the field of biosensors. This year Prof. Guiseppi-Elie and research associate Dr. Sean Brahim, again with University of Texas collaborator, Raymond Baughman, with contributions from Case Western Reserve University Engineering Professor, Dr. Gary Wnek, have submitted yet another paper, “Carbon Nanotube Modified Electrodes for the Direct Bioelectrochemistry of Pseudoazurin”, extending this work to the bioelectrochemistry of the metalloprotein pseudoazurin. The latter paper has now been accepted for publication in the inaugural issue of the recently formed journal, NanoBiotechnology, In both these papers the authors demonstrate how these novel carbonaceous materials can be made to shuttle electrons to and from the redox centers of these enzymes, thereby reducing the dependence on mediators such as oxygen and other small redox molecules.

Design of a Subcutaneous Implantable Biochip for Monitoring of Glucose and Lactate:

In a similarly titled manuscript accepted for publication in IEEE Sensors Journal (2005), Prof. Guiseppi-Elie and C3B researchers Dr. Sean Brahim and Gymama Slaughter, in collaboration with VCURES Associate Director, Dr. Kevin Ward, describe the design and fabrication of a miniature (2 mm x 4 mm) electrode coated with a thin, sensing membrane that responds selectively to both glucose and lactate. The impetus for this research comes from the fact that at present there is no clinically reliable in vivo biosensor that can be used for accurate, continuous, long-term sensing (> 6 months) and reporting of important physiologic metabolites such as glucose and lactate. The need for such a device is absolutely critical for the successful development of truly ‘closed-loop’ systems for treating epidemics such as diabetes and also in combat and civilian casualty care. The electrode fabrication process includes standard microlithographic techniques The biorecognition layer of the working electrodes was fabricated from an ca. 1.0 mm thick composite membrane of principally tetraethylene glycol (TEGDA) cross-linked poly(2-hydroxyethyl methacrylate) that also contained a derivatized polypyrrole monomer and a biomimetic methacrylate component with pendant phosphorylcholine groups. These two additional monomer components were introduced to provide interference screening and in vivo biocompatibility respectively; two major issues that have plagued the successful development of implantable biosensors in the past. Analyte specificity was achieved by immobilizing the appropriate enzyme(s) within the hydrogel membranes. In vitro analytical characterization of the biochip has demonstrated good linear dynamic response ranges, sensitivities, and response times towards both glucose and lactate in simulated physiologic media. In vivo studies in rats are currently in progress. This work is supported by a grant (PR023081) from the US Department of Defense (DoDPRMRP).

Two important papers on robust engineering of DNA Microarrays for clinical applications:

In two recently published papers, Prof. Guiseppi-Elie and his students along with VCU Medical Center collaborators describe the design and evaluation of a 20,000 spot human oligonucleotide microarray. The first of these papers address the selection of immobilization chemistries for the attachment of oligonucleotide sequences to DNA microarrays. [Scott Taylor1, Stephanie Smith1, Brad Windle2, and Anthony Guiseppi-Elie1,3,* (2003) Impact of surface chemistry and blocking strategies on DNA microarrays, Nucleic Acids Research Vol. 31, No. 16]. The second paper investigates and reports on the use of experimental design techniques to evaluate the quality control and assurance impact of potentially variable factors on gene expression results derived from the oligonucleotide microarray. [Scott Taylor1 Kellie J. Archer2, and Anthony Guiseppi-Elie1,3,4,* (Submitted) A Factorial Study of the Influence of Production and Protocol Variables on Gene Expression Data From Spotted Oligonucleotide Microarrays Biotechniques (submitted). Both papers are aimed at designing developing clinically hardened microarray technology

Biosensors Presentations

1. ‘Development of a Temporary Implantable Biochip for Monitoring of Glucose and Lactate During Hemorrhage’, Anthony Guiseppi-Elie, Sean Brahim, Sheena Abraham, Gymama Slaughter, Robert Diegelmann, Luciana Torres, R. Wayne Barbee and Kevin Ward, Advanced Technology Applications for Combat Casualty Care 2003 Conference, Tradewinds Island Beach Resort, St. Pete Beach, St, Petersberg, Florida, USA, August 16-18, 2004.

2. ‘Carbon Nanotube Modified Electrodes for the Bioelectrochemistry of Redox Enzymes’, Sean Brahim, Chenghong Lei, Gary Wnek, Ray Baughman, and Anthony Guiseppi-Elie, 2004 Virginia Nanotechnology Showcase: Exploring Research and Commercialization in Nanomanufacturing, Omni Hotel, Charlottesville, Virginia, USA June 22 & 23, 2004.

3. ‘Development of a Temporary Implantable Biochip for Monitoring of Glucose and Lactate During Hemorrhage’, Anthony Guiseppi-Elie, Sean Brahim, Sheena Abraham, Gymama Slaughter, Robert Diegelmann, Luciana Torres, R. Wayne Barbee and Kevin Ward, US Department of Defense Peer Reviewed Medical Research Program (PRMRP) Investigators Meeting and Military Health Research Forum, Caribe Hilton, San Juan, Puerto Rico, April 25 – 28, 2004.

4. ‘Carbon Nanotube Modified Electrodes for the Bioelectrochemistry of Redox Enzymes’, Sean Brahim, Chenghong Lei, Gary Wnek, Ray Baughman, and Anthony Guiseppi-Elie, 2004 Virginia Commonwealth University Nanofiber Symposium, Virginia Biotechnology Research Park, Richmond, Virginia, USA April 16, 2004.

5. ‘Feasibility Studies in Development of a Temporary Implantable Lactate Sensor Biochip for Monitoring During Hemorrhage’, Anthony Guiseppi-Elie, Sean Brahim, Sheena Abraham, Gymama Slaughter, Felix Miranda, Rainee Simons, Luciana Torres, Robert Diegelmann, Wayne Barbee, and Kevin Ward, Advanced Technology Applications for Combat Casualty Care 2003 Conference, Tradewinds Island Beach Resort, St. Pete Beach, St, Petersberg, Florida, USA, August 18-20, 2003.

6. ‘Characterization of Electroconductive PPy-p(HEMA) Composite Hydrogels For Sensing Applications’, Sean Brahim and Anthony Guiseppi-Elie, 81^st Annual Meeting of the Virginia Academy of Science, University of Virginia, Charlottesville, Virginia, USA, May 29, 2003.

7. ‘Design of a Subcutaneous Implantable Biochip’, Anthony Guiseppi-Elie, Sean Brahim and Kevin Ward, C3B Spring 2003 External Advisory Board Meeting, Virginia Commonwealth University, Richmond, Virginia, USA, May 8, 2003.

8. ‘Comparative Evaluation of Bio-smart Hydrogels for Biosensors’, Gymama Slaughter, Sean Brahim and Anthony Guiseppi-Elie, 16^th Annual Meeting of the Mid-Atlantic Biochemical Engineering Consortium (MABEC) 2003, University of Maryland, College Park, Maryland, USA, March 14, 2003.

9. ‘Bio-smart Composite Hydrogel Polymers: Integrated Biosensing and Controlled Drug Release’, Sean Brahim and Anthony Guiseppi-Elie, NIH Bioengineering Consortium (BECON) 2002: Sensors for Biological research and Medicine, Natcher Conference Center, National Institutes of Health, Bethesda, Maryland, USA; June 24-25, 2002.

10. ‘Bio-smart Composite Hydrogel Polymers: Integrated Biosensing and Controlled Drug Release’, Sean Brahim and Anthony Guiseppi-Elie, Bioinformatics & Phamacogenomics Symposium 2002, Virginia Bioinformatics Consortium, Omni Richmond Hotel, Richmond, Virginia, USA; June 12-13, 2002.

11. ‘Bio-smart Hydrogels: Co-joined Molecular Recognition and Signal Transduction in Biosensor Fabrication and Drug Delivery’, Sean Brahim and Anthony Guiseppi-Elie, 80^th Annual Meeting of the Virginia Academy of Science, Hampton University, Hampton, Virginia, USA; May 21-24, 2002.

Biochips Presentations

1. Mid-Atlantic Bioengineering Conference (MABEC) 2002

· Detection of DNA Hybridization by Impedance and Development of Impedimetric DNA Microarray for Clinical Classification of Brain Tumors G. Scott Taylor, Stephanie Smith, Marine Gheorghe, Derk Bemeleit, Dietmar Blohm, Oliver Bogler, William Broaddus, and Anthony Guiseppi-Elie.

2. Mid-Atlantic Bioengineering Conference (MABEC) 2003