NC1194: Nanotechnology and Biosensors
Annual/Termination Reports (SAES-422): [10/15/2012]
Date of Annual Report: 10/15/2012
Report Information:
Participants:
- Alocilja, Evangelyn (alocilja@msu.edu) Michigan State University
- Bhalerao, Kaustubh (bhalerao@illinois.edu) University of Illinois
- Bralts, Vincent (bralts@purdue.edu) - Purdue University
- Rastogi, Shiva (srastogi@uidaho.edu) University of Idaho
- Su, Winston (wsu1546@gmail.com) - University of Hawaii
- Takhistov, Paul (takhistov@aesop.rutgers.edu) - Rutgers University
- Yoon, Jeon-Yeol (jyyoon@email.arizona.edu) University of Arizona
- Yu, Chenxu (chenxuyu@iastate.edu) - Iowa State University.
Brief Summary of Minutes of Annual Meeting:
The meeting was held on July 5-6, 2012 at the Kellogg Hotel and Conference Center, Room 105A, Michigan State University, 219 S. Harrison Rd., East Lansing, MI 48824.First Day, July 5, 2012
At 8:15am on Thursday, July 5, 2012, the meeting was called to order by Dr. Evangelyn Alocilja, 2012 chair of the multi-state group NC-1194, Nanotechnology and Biosensors. The meeting started with introduction of the participants. The group was welcomed to MSU by the following MSU administrators: Dr. Steve Pueppke, Associate Vice President for Research and Graduate Studies, and Director, AgBioResearch; Dr. Satish Udpa, Dean, College of Engineering; and Dr. Ajit Srivastava, Chair, Department of Biosystems and Agricultural Engineering. Following the welcome, the NC-1194 members presented their progress reports. Questions and discussions of the reports followed each presentation. Discussion covered significance, intellectual merit, and broader impact. Titles of participant presentations are given in Table 1.
After the institutional reports, the group discussed potential multi-institutional collaborations. The group looked at the following opportunities: USDA-NIFA RFPs, Nanotechnology Signature Initiative entitled Nanotechnology for Sensors and Sensors for Nanotechnology, NSF engineering research centers, international collaboration, and instructional collaborations, such as the potential for sharing resources, course materials, and teaching time. One opportunity for collaboration is joint publication. Dr. Chenxu Yu is guest editing a special issue for the open access journal Biosensors, with a title Nanomaterials for biodetection and drug delivery. He invited the group to submit research papers for this issue. Deadline for paper submission is January 31, 2013.
At 3pm, Dr. Alocilja presented opportunities for technology commercialization and challenged the group to think on how to move their technologies from the lab to the field through effective mechanisms, private-public partnerships, and relevant applications.
At 4pm, Dr. Elliot Ryser (MSU) presented the objectives and mission of the multi-state Enhancing Microbial Food Safety by Risk Analysis. The group discussed with Dr. Ryser potential collaboration between the two multi-state groups.
The group had a working dinner to elect the next set of officers and identify the location of the next meeting. The elected officers for 2013 are:
Chair: Daniel Jenkins
Vice Chair: Jeong-Yeol Yoon
Secretary: Kaustubh Bhalerao
Next meeting: The 2013 annual meeting will be hosted by the University of Hawaii and to be coordinated by Dr. Daniel Jenkins.
Second Day, July 6, 2012
Dr. Alocilja called to order the meeting for the second day, Friday, July 6, 2012. Dr. Douglas Buhler, Interim Dean, College of Agriculture and Natural Resources, was there to welcome the group. At 8:30am, the group drove to the Cyclotron Building to get a 9am tour of the Cyclotron Facility and the Facility for Rare Isotope Beams (FRIB), a new national user facility for nuclear science which is funded by the Department of Energy Office of Science and the state of Michigan.
At 10:30am, the group visited the Nano-Biosensors Lab in the Department of Biosystems and Agricultural Engineering.
The meeting was adjourned at 12:00 noon.
Table 1. Institutions, participants, and title of presentations. Rutgers University, Paul Takhistov, Nanoparticles in Food: biosensing, nanostructured materials and transport
University of Illinois at Urbana-Champaign, Kaustubh Bhalerao, Synthetic biology and bionanotechnology
Iowa State University, Chenxu Yu, NanoDEP/SERS-LSPR sensor platform, nano-enabled bactericidal film for food processing, and other technologies
University of Idaho, Shiva K. Rastogi, DNA Detection on Lateral Flow Test Strips: Enhanced Signal Sensitivity Using LNA-conjugated Gold Nanoparticles
University of Hawaii, Wei Wen Su, Tunable nano-oleosomes: biosynthesis, characterization, and applications
University of Arizona, Jeong-Yeol Yoon, Development of handheld cell-phone based biosensor and microfluidic devices
Michigan State University, Evangelyn Alocilja, Nano-Biosensors for Global Health, Bio-defense, Food Safety, and Water Quality
Accomplishments:
This is a progress report for the period October 1, 2011-September 30, 2012 by participating institutions covering various states. As a reminder, the objectives of this project are: 1. Develop new technologies for characterizing fundamental nanoscale processes 2. Construct and characterize self-assembled nanostructures 3. Develop devices and systems incorporating microfabrication and nanotechnology 4. Develop a framework for economic, environmental and health risk assessment for nanotechnologies applied to food, agriculture and biological systems 5. Produce education and outreach materials on nanofabrication, sensing, systems integration and application risk assessment Progress reports are presented by state. The findings have been disseminated to the scientific community via seminars, national/international conferences, manuscripts, and web sites.Arizona (University of Arizona) Accomplishments from the University of Arizona were presented at the meeting by Dr. Jeong-Yeol Yoon (Table 1). For details, please refer to http://biosensors.abe.arizona.edu/index.html.
Idaho (University of Idaho) Accomplishments from the University of Idaho were presented at the meeting by Dr. Shiva Rastogi (Table 1). For details, please refer to http://www.uidaho.edu/cals/news/feature/rastogi.
Illinois (University of Illinois at Urbana-Champaign) Accomplishments from the University of Illinois at Urbana-Champaign were presented at the meeting by Dr. Kaustubh Bhalerao (Table 1). For details, please refer to http://abe-bhaleraolab.age.uiuc.edu/members/kbhalerao/
Indiana (Purdue University) Output. In the reporting year, principal investigators at Purdue University made advances in the area of nanotechnology and biosensors. These advances include novel and improved technologies for both intracellular and extracellular measurements of dynamic biomolecule events within cells. Fundamental understanding and design of material properties and behavior at the nanoscale fueled these technologies. These properties include electrochemical, optical, and biological behavior of carbon nanotubes, nanostructures platinum, mesoporous silica, platinum nanoparticles, and gold nanoparticles and probes. These physiological sensing technologies have a wide range of applications and impact in areas including environmental toxicology and monitoring, metabolic disease including diabetes, cancer, water monitoring and regeneration, and food.
Iowa (Iowa State University) Accomplishments from Iowa State University were presented at the meeting by Dr. Chenxu Yu (Table 1). For details, please refer to http://www.abe.iastate.edu/abe-department/directory/chenxu-yu/. A written report is also presented below. Output. The activities during this period included: (1) further development of dual-recognition Raman sensing scheme for single step pathogen detection in a lab-in-a-tube platform, detection limit is improved to 100 CFU/mL, with a multiplex target identification scheme. (2) The development of nanophotocatalyst for pretreatment of biomass to improve its digestibility for biofuel/biorenewables production. (3) The development of Raman spectroscopic imaging-based characterization of glaucomatous retinal tissues. The Raman imaging technique may serve as the basis for a method to diagnose glaucoma at an early stage, which will greatly benefit glaucoma patients. (4) The development of a nano-functionalized antimicrobial biodegradable coating for food safety control. (5) The development of a rapid UV spectroscopic detector for monitoring trace amount of egg yolk contamination in egg white. These findings were presented in several peer reviewed conferences and three journal articles. Impact. The spectroscopic sensing techniques developed in this project are highly accurate, they have the potential to meet the needs in various disciplines for high-accuracy target sensing, including early diagnosis of diseases (i.e., glaucoma), and rapid screening for foodborne pathogens. The utilization of nanophotocatalyst to pretreat biomass with reduced water and chemical usage, which will make biofuel and biorenewable production more environmentally friendly and less costly. The development of a nano-functionalized antimicrobial biodegradable coating for food safety control utilizes biodegradable films made from soy and corn proteins, and functionalizes them with antimicrobial nanoparticles to improve their mechanical strength and to impart in the film antimicrobial functionality. The goal is to create cheap coating materials that are antimicrobial and can be used in food processing/food handling places to improve food safety, such as fresh produce and meat. The UV spectroscopic sensor for egg yolk contamination in egg white addresses an important industrial need. It can detect 0.001% yolk contamination in egg white in less than 5 seconds.
Hawaii (University Hawaii) Accomplishments from the University Hawaii were presented at the meeting by Dr. Wei Wen Su (Table 1). For details, please refer to http://www2.hawaii.edu/~wsu/. A written report is also presented below. Output. Research for the project has focused on the development of technologies enabling rapid gene-based agricultural diagnostics directly in the field. Activities/outputs include: (1) A new molecular probe technology Assimilating Probes was developed which was the first to enable real-time quantitative detection using an isothermal gene based amplification LAMP, and which can be used for multiplexed detection of multiple sequences or the incorporation of an internal positive control to validate test performance. Primer probe sets have been developed for several pathogens, including Ralstonia solanacearum and Salmonella enterica. (2) A single-test handheld device was developed for rapid field detection using LAMP and assimilating probes. Devices, interfaced by Bluetooth with a tablet or smart-phone, were demonstrated in the field at a remote village in Guatemala for the detection of select agent race 3 biovar 2 strains of the bacterial wilt pathogen Ralstonia solanacearum. (3) A multiplexed LAMP/ Assimilating Probe assay was developed to classify race 3 biovar 2 strains of Ralstonia solanacearum, allowing discrimination of these quarantine strains from more ubiquitous and less harmful strains of the pathogen. (4) A second generation handheld device was developed capable of running 8 simultaneous test reactions, with two optical channels for detection of two specific sequences in each reaction. This design is currently being refined into a commercial-ready device suitable for sale to researchers, food producers and processors. (5) Work has been initiated on a new method for screening large volumes of agricultural materials (rinsates, irrigation water, soil, plant and animal tissues) to effectively isolate disperse zero-tolerance pathogens such as Salmonella and lethal E. coli strains. Results have been disseminated to scientific communities and to the public through a variety of publications and conference presentations. Impact. The technologies developed in this research are intended to allow rapid detection of agricultural and food-borne pathogens to improve food security and safety. Handheld rapid detection has been identified as a critical need by the USDA for enhancing food safety and implementing effective surveillance programs to exclude harmful pathogens from agricultural settings. We have initiated collaborations with a number of USDA-ARS research groups to adapt our technologies for serious food-pathogens, devastating quarantine pathogens, and other pest organisms to facilitate more effective/ economical pest management. In the 2011 annual project meeting, commercialization of sensor technologies was identified as being a critical activity as the technologies themselves are rapidly reaching maturity. As such, we have focused significant efforts in commercializing inexpensive devices and gene-based test kits that can be a powerful tool for food growers and processors.
Michigan (Michigan State University) Accomplishments from Michigan State University were presented at the meeting by Dr. Evangelyn Alocilja (Table 1). For details, please refer to http://www.egr.msu.edu/~alocilja/. A written report is also presented below. Output. Research accomplishments at Michigan State University include the development of a green chemistry synthesis of gold nanoparticles, development of nanoparticle-based biosensors, validation of these biosensors in various food matrices, and effective immunomagnetic extraction techniques. Specifically, we have developed biosensors based on biobarcode, electrospun nanofiber, nonwoven fibers, polyaniline label, and copolymer mass amplification. We have also developed a method for directly detecting genomic DNA from Salmonella enteritidis without PCR amplification. Impact. In January 2012, Dr. Alociljas nano-biosensor technologies have been licensed to nanoRETE, Inc., a start-up company established to commercialize her technologies. The company is backed by Michigan Accelerator Fund I, a venture capital investment group based in Grand Rapids, Michigan. Dr. Alocilja is the companys founder and Chief Scientific Officer. The company is located in Lansing, MI. With 4 full time scientists and 3 part-time researchers and administrators, the company is already contributing to job creation in Michigan.
Minnesota (University Minnesota) Output. The main problems in the dental implant industry are: (1) formation of biofilm around the implant a condition known as peri-implantitis, and (2) inadequate bone formation around the implant lack of osseointegration. Therefore, developing an implant to overcome these problems is of significant interest. Chitosan has been reported to have good biocompatibility and also anti-bacterial activity. An osseo-inductive recombinant biopolymer (P-HAP), derived from the protein statherin has been reported to induce bone formation. We have deployed these two materials to build assembled layer-by-layer (LbL) coatings on titanium by exploiting their opposite charges in appropriate conditions. The LbL coatings were characterized by contact angle measurement, FTIR and AFM. The substantial difference in water contact angle between alternate layers, the representative peaks in FTIR & XPS, and the changes in topography between surfaces with different number of bi-layers observed in AFM indicated that we successfully built-up the chitosan/P-HAP LbL. The LbL samples showed increased amount of biomineralization in osteogenic media (OS) supplemented with CaCl2, which is attributed to the well-established preferential interaction of statherin with mineral in teeth. The osteoblast cells line (MC3T3) adhered well to the constructed bi-layers, but did not quite differentiate well on the bi-layers. Since the biomineralization increased with the layer buildup, these biomineralized samples can be further explored to assess their biological activity - cell adhesion, proliferation and differentiation of osteoblasts and antimicrobial - inhibition of attachment and growth of oral pathogens in vitro potential.
New Jersey (Rutgers University) Accomplishments from Rutgers University were presented at the meeting by Dr. Paul Takhistov (Table 1). For details, please refer to http://foodsci.rutgers.edu/takhistov/.
New York (Cornell University) Output. We have worked on the development of a number of different platforms over the course of the last year. This includes microfluidic channels for the isolation and amplification of RNA molecules of the protozoan parasite Cryptosporidium parvum; transfer of the technology to a commercial microfluidic system in collaboration with Rheonix, Inc., by investigating the possibility of using biofunctional electrospun nanofibers directly in microfluidic channels; by integrating a pre-concentration step on-chip; and by investigating novel signaling strategies for lab-on-a-chip devices based on electrochemiluminescence. Impact. We have developed a minipotentiostat that can control a 3-electrode system and can be used for electrochemiluminescence detection. We have determined that nanovesicles and cells can be specifically captured on appropriate nanofiber surfaces and can be released using pH shifts. We have demonstrated that pre-concentration prior to detection on-chip can lower the limit of detection by more than one order of magnitude. Over the past decade, rapid point-of-care (POC) tests have emerged as one of the largest and fastest growing segments of the human in vitro diagnostic testing market and can also be found in food safety applications. Due to their relative ease, most such tests rely upon immunologic methods. A common testing format employs specific antibodies that recognize and interact with the analytes of interest in a manner that will also induce the generation of a discernable color within a lateral flow device. Such tests are frequently designed to yield qualitative (i.e., yes or no) results rather than quantitative tests. Despite the added information that can be obtained via molecular detection methods, similar rapid tests employing gene detection methods have not yet found widespread use, which is primarily due to the more complex and time-consuming nature of sample preparation methods as well as the more technically complicated gene amplification techniques. The approach developed by us will greatly ameliorate the situation since it combines a quantitative approach with the highly selective nucleic acid recognition of pathogenic organisms and simplifies the amplification techniques. Thus, pathogens will be detectable and quantifiable at significantly lower costs, and tests will be more reliable since fewer assay steps are needed.
South Carolina (Clemson University) Output. Adherence is among the earliest event in many bacterial infections. These interactions are required for extracellular colonization and/or internalization. This colonization is usually mediated by bacterial adhesins on the surface of bacteria that recognize and bind to specific receptor moieties of host cells. This adhesin-receptor binding event could activate cascades of signal transduction important in the pathogenic process and host-defense. Understanding of this adhesin-receptor interaction is of great value in order to develop effective prevention, detection, diagnostic, and treatment methods. Nanotechnology offers new opportunities for the nanoscale investigation of adhesin-receptor interactions. We envision that adhesin specific nanoparticles and biosensors will break new ground in study of pathogen-host interactions, drug discovery, drug delivery, and biosensing. To achieve such goals, the fundamentals of adhesin-receptor interactions must be examined. In this present work, the PI and his team have functionalized nanoparticles with receptor molecules recognized by specific bacterial adhesins to demonstrate that such nanoparticles could be utilized to eliminate pathogens from host without the need for antibiotics that many pathogens are now resistant to. The PI and his team have also utilized the same principle to develop highly sensitive biosensors capable of detecting the targeted pathogens in low number. The PI has received a resource grant from the Consortium for Functional Glycomics allowing the PI to tap into their carbohydrate resources enabling the PI to functionalize various nanomaterials for the proposed studies. The PI and his team have also fabricated several nanomaterials/devices to study nanoscale phenomena for applications in pathogen detection/inactivation and risk assessment. Impact. The use of carbohydrates as capturing agents in biosensor development represents a more specific, stable, and economical approach than the conventional immunoassays or PCR detection methods that are more costly and incapable of differentiating non-viable from viable targets.
Wisconsin (University of Wisconsin) Output. We are investigating how a fundamental nanotechnological phenomenon plasmonic effect of in situ synthesized gold nanoparticles (AuNPs) can be used to develop a novel system to provide visible indication of thermal history of foods, which will signal the need for prompt and timely food safety intervention. Formation of AuNPs can mimic temperature-driven changes in bioproducts by using functional biomolecules (proteins/polysaccharides) as mediate materials, because the specific characteristics of the biomolecules dictate the formation of AuNPs. Gelatin is an edible protein derived from collagen. Due to its low cost and unique ability to form thermo-reversible hydrogel, gelatin is used in a myriad of practical applications including as a food and as stabilizing agent in foods. Gelatin has been employed for the synthesis of metal NPs. The outstanding transparency of gelatin gels is also highly valued, especially by the photographic film industry for colorimetric applications. Thus, gelatin is an excellent material for both mediating the formation of AuNPs and representing biomaterials. We are studying AuNPs synthesis in the presence of gelatin as a potential system for food material thermal history by investigating the effect of temperature and temperature history on the AuNP synthesis. Impact. Our system will help ensure the safety and security of our food chain before foods contaminated with pathogenic organisms and/or toxins are distributed and consumed.
Work plan for the next year: Research by each investigator at each participating institution will continue as funding becomes available. New techniques will be developed for characterizing fundamental nanoscale processes. Self-assembled nanostructures will be designed and characterized. New biosensor devices and systems will be developed and will continue to incorporate microfabrication and nanotechnology. Economic frameworks and environmental and health risk assessments of nanomaterials as applied to food, agriculture and biological systems will become a priority. And finally, educational and outreach materials on nanofabrication, sensing, systems integration and application risk assessment will be developed.
Impact Statements:
- The technologies developed in this multi-state collaboration are designed to improve food safety, water quality, agricultural production, and health. Handheld biosensor devices utilizing nanomaterials and techniques are the major contributions in this report. Furthermore, transfer of some of the technologies to the private sector has generated jobs and will contribute to the economic progress of the country and respective states.