Source: UNIV OF MASSACHUSETTS submitted to
PARTNERSHIP: DEVELOPMENT AND EVALUATION OF LOW-COST, EASILY DEPLOYABLE MOLECULARLY IMPRINTED POLYMER NANOPARTICLES FOR AGRICULTURAL VIRUSES AND TOXINS OF CONCERN
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
ACTIVE
Funding Source
AFRI COMPETITIVE GRANT
Reporting Frequency
Annual
Accession No.
1028014
Grant No.
2022-67021-36408
Cumulative Award Amt.
$743,868.00
Proposal No.
2021-08570
Multistate No.
(N/A)
Project Start Date
Feb 1, 2022
Project End Date
Jan 31, 2026
Grant Year
2022
Program Code
[A1511]- Agriculture Systems and Technology: Nanotechnology for Agricultural and Food Systems
Recipient Organization
UNIV OF MASSACHUSETTS
(N/A)
AMHERST,MA 01003
Performing Department
Dept: Food Sciences
Non Technical Summary
Foodborne illness causes considerable damage to public health and causes considerable economic loss in the United States and globally. The leading cause of foodborne infections in the US and globally is human norovirus, which causes millions of illnesses, results in hundreds of thousands of deaths, and losses in billions of dollars annually. These viruses can survive on surfaces for weeks, are resistant to most commonly used disinfectants, and require just a handful of particles to make someone sick.Mycotoxins are toxins that certain fungi can produce that cause illness in humans and agricultural animals. Although illness from mycotoxins does not immediately result in direct symptoms, these toxins can cause severe damage to peoples' liver and kidneys over time if consumed, making them even harder to prevent and control. Mycotoxins occur when certain fungi grow on foods that are eventually consumed, and the foods most commonly associated with these toxins include grains, legumes, and other plant-based products. Further, the conditions that climate change is causing have potential to further promote growth of these fungi and subsequent occurrence of mycotoxins in foods. This potential for higher presence of these toxins in foods due to climate change, as well as an increasing trend (and need for) of consumption of plant-based foods, makes mycotoxins and emerging existential public health threat. Despite their importance, routine and comprehensive testing for noroviruses and mycotoxins in foods is still fairly limited as a practice due to the lack of rapid, portable, and inexpensive testing methods and protocols; thus making controlling their consumption more difficult.A number of challenges exist in testing for noroviruses and mycotoxins in foods. One of which is that both are very diverse in the different types/variants of these contaminants that exist. One must be able to detect the broad class, while also being able to determine which type is detected. Further, many testing techniques either take a long time (limiting application), are not suitable for use in fields (require central lab testing), can be easily inhibited by other compounds found in foods, and are too costly to be realistically utilized for noroviruses and mycotoxins.Molecularly imprinted polymer nanoparticles (nanoMIPs), are an emerging technology that offers the potential for the development of inexpensive testing technologies that are also able to withstand harsh conditions (in-field). The application of nanoMIPs for norovirus and mycotoxin testing has not been investigated, and this project plans to develop this promising technology for the development of tests that are portable, inexpensive, and have potential to be utilized to target specific types of noroviruses and mycotoxins, thus enabling the ability to better detect noroviruses and mycotoxins in foods before they are served to people. The project will also evaluate the ability of the nanoMIP tests to withstand and perform in foods without the need to complicated manipulation that will add time to result and limit their application at the point of production or service. Overall, the development of such tests for noroviruses and mycotoxins has potential to reduce the considerable public health and economic loss that noroviruses and mycotoxins provides, as well as lead to knowledge that will aid development of nanoMIP sensors for other viruses and toxins of concern.This project also involves the establishment of an international partnership between two globally-leading research institutions, University of Massachusetts Amherst and Newcastle University. The Moore and Gibbons labs in the Department of Food Science at the University of Massachusetts are respective experts in foodborne viruses and fungi, respectively. The Peeters lab in the School of Engineering at Newcastle University has expertise in nanoMIPs and the development of rapid tests. Further, the Department of Food Science is ranked among the top departments in food science research in the US and globally, while Newcastle University is recognized as among the global leaders in the development of sensing technologies. This partnership is not only ideal for development of these novel testing methods for noroviruses and mycotoxins, but will also result in forming the foundation of a partnership that is anticipated to result in future scientific research. The partnership will also involve exchanges and training of the labs involved, and will also include additional activities to solidify collaboration between these two world-leading institutions.
Animal Health Component
60%
Research Effort Categories
Basic
30%
Applied
60%
Developmental
10%
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
7124030110150%
7124020115050%
Knowledge Area
712 - Protect Food from Contamination by Pathogenic Microorganisms, Parasites, and Naturally Occurring Toxins;

Subject Of Investigation
4020 - Fungi; 4030 - Viruses;

Field Of Science
1101 - Virology; 1150 - Toxicology;
Goals / Objectives
This projectseeks to develop and evaluate the potential of molecularly imprinted polymer nanoparticles (NanoMIPs) to serve as recognition elements for food and agricultural contaminants. We have chosen two targets of serious relevance to foodborne contamination, norovirus and mycotoxins that also serve as model targets in that they both are of smaller size, have a diversity of structures, and are more predominantly found in different food matrices. Further, the project will integrate these nanoMIPs into inexpensive, portable electrochemical sensors. We additionally hope to evaluate the degree to which nanoMIPs and nanoMIP sensors generated against these targets are able to withstand crudely processed food samples containing target contaminants, as well as the degree to which these sensors are capable of discriminating different variants/strains of noroviruses and mycotoxins. Additionally, the ability of the nanoMIP sensors to stand up to harsher field conditions will be evaluated. The project aims to result in a number (at least 4) of publications as well as posters and scientific presentations as a result of this work.One of the other major goals of the project is to establish a partnership between the University of Massachusetts, Amherst and Newcastle University. The project will include an exchange where PI and selected students/postdoc will conduct an exchange that will involve guest seminars/networking for future collaboration by the PIs with each other's respective institutions, as well as an expertise exchange between labs from the two institutions. The Moore lab (UMass) has expertise in applied microbiology, virology, and detection of pathogens in food and agricultural samples. The Peeters lab (Newcastle) has expertise in nanoMIP development, biosensor fabrication, and electrochemical sensing evaluation. The Gibbons lab (UMass) has expertise in fungal pathogens and their toxins. Students/postdocs from these two labs will spend at least one week in each other's' labs learning the techniques of each other's' labs. The Department of Food Science at the University of Massachusetts, Amherst is a global leader in food science, recently ranked first in the U.S. and second in the world in food science by U.S. News & World Report. Newcastle University is a noted global leader in electrochemical sensing and sensor engineering. Built in seminar and faculty meetings that will coincide with the visit will further promote collaboration. An additional goal of the project is to utilize what is learned from the evaluation of the nanoMIP technology into intellectual property and potential future devices for the agricultural market, as the devices themselves are anticipated to be inexpensive enough to be realistically utilized by the food production industry.The specific objectives of this project are:Objective 1. Development and evaluation of molecularly imprinted polymer nanoparticles (nanoMIPs) for detection of noroviruses and integration into an inexpensive electrochemical sensor.Objective 2. Development and evaluation of molecularly imprinted polymer nanoparticles for the detection of mycotoxins and integration into an inexpensive electrochemical sensor.Objective 3. Evaluation of the ability of electrochemical nanoMIP sensors to withstand common inhibitory substances as well as performance in crudely processed, real food samples in in-field conditions.
Project Methods
The methods of the project will include the specific activities outlined in the revised narrative submitted as a part of the response to panel comments. Specifically, numerous conditions involved in nanoMIP development and different aspects of the different selected targets will be executed. The performance of nanoMIP candidates will be evaluated by determination of binding affinity of the nanoMIPs for both the specific target norovirus/mycotoxin target strain/variant as well as others of varying degrees of relatedness (specificity). The ability of the nanoMIPs to withstand harsher field conditions (higher temperature) will also be determined in part by affinity and sensitivity for target when integrated into sensors. The ability of the nanoMIPs when incorporated into sensors to detect norovirus and mycotoxin targets will be evaluated based on sensitivity (limit of detection) and specificity (reactivity with different strains/variants). The ability of the nanoMIP sensors to withstand inhibitory components found in foods (both purified known inhibitors and crudely processed food samples) will also include sensitivity.

Progress 02/01/24 to 01/31/25

Outputs
Target Audience:The target audience reached by the work in this reporting period consists primarily of fellow academics and researchers in the fields of detection/analytical biochemistry, microbiology, materials and bioengineering, and food safety. Secondary audiences reached in the different presentations/communications/publications involves members of the food/food safety industry, government, and academia. In particular, our data related to the potential of this technology to rapidly detect viruses as well as more fundamental developments in MIP technology to detect antibiotics in foods would be of interest to the broader infectious disease, polymer science, and food safety communities as a whole--including those who are focused on pathogen/contaminant control from a clinical perspective. Changes/Problems:Co-PI Peeters moved institution from Newcastle University to the University of Manchester during this period, which caused a major pause in the progress of the research for this award, as the Peeters lab generate the nanoMIPs. In good news, co-PI Peeters also gave birth to a baby and took a half year maternity leave, though her lab was settled into U Manchester and able to conduct research. What opportunities for training and professional development has the project provided?This was a very successful year for professional training activities. We had a number of poster presentations and technical talks both within the United States and England, as well as internationally outside of the PIs' home countries (Latvia, Italy, Belgium, Canada). One PhD student defended her PhD during this period, and one Postdoc obtained a research faculty position at Pukyong National University in Busan, Korea. Further, 2 other graduate students had the opportunity to present posters, and two undergraduates received direct training from the senior graduate students/postdoc, expanding mentorship skills. Graduate students also delivered some guest lectures about detection in foods to different undergraduate/graduate lectures (5 guest lectures for 4 classes). Additionally, both PIs Moore and Peeters have aided and supported applications for different awards and fellowships for students and postdoc funded by the project. This has resulted in the students associated with this project winning multiple awards, some of which include: an ASM Peggy Cotter Young Scientist Travel Award (postdoc), Alliance to Stop Foodborne Illness 40 Food Safety Professionals Under 40 (senior grad student--extremely prestigious), New England IFT Graduate Student Scholarship, UMass Food Science Manley Graduate Scholarship (top award for department PhD student), and Phi Tau Sigma Student Achievement Scholarship. Additionally, the PIs of the project were given awards, including: Promotion to Associate Professor (Moore) and Full Professor and Chair in Engineering Biology (co-PI Peeters); Eric A. Decker Scholar (Inaugural, Moore), IFT Outstanding Young Scientist Award in honor of Samuel Cate Prescott (Moore), and Foods Young Investigator Award (Moore).? How have the results been disseminated to communities of interest?As mentioned above, the results and accomplishments have all been presented through numerous products of traditional means, like a publication, textbook chapter, posters, technical talks, and invited talks (above). Although not directly about the specific project, the PIs' expertise as a part of awards like this has been recognized by appearing as experts in the popular press, such as: 1. New York Times: "Are Eggs Safe to Eat as Bird Flu Spreads?" February 2, 2025. 2. Food Technology: "Cracking the Code on Research Funding." Cover Story. December 2024/January 2025 Issue. 3. The Montague Reporter: "Area Farmers, Birders on Alert As Avian Flu Outbreak Worsens." January 2, 2025. 4. New York Times: "How Worried Should We Be About Bird Flu?" December 18, 2024. 5. Brain Food--The Official Blog of IFT: "The Cost of Discovery." Interview Feature. December 13. 6. New York Times: "Is Milk Safe? Your Questions About Bird Flu and Dairy, Answered." December 6, 2024. 7. USDA NIFA Press Release: "USDA NIFA Invests $14 Million to Further Food Safety Research and Outreach." November 20, 2024. 8. Parents.com: "Why Have There Been So Many Food Recalls Lately? Experts Explain." October 31, 2024. 9. New York Times: "Can Drinking Alcohol Prevent Food Poisoning?" September 16, 2024. 10. Food Processing (Australia): "Developing methods to streamline foodborne virus detection." August 12, 2024. 11. UMass CNS: "Matthew Moore Receives $650K Grant to Detect Viruses in Food." August 12, 2024. 12. Boston Globe: "Six things to know about bird flu in milk." May 20, 2024. 13. UMass News: "UMass virologist Matthew Moore wins outstanding young scientist award." May 8, 2024. 14. Institute of Food Technologists Press Release: "Institute of Food Technologists Honors Top Leaders in Science of Food Community with 2024 Fellows, Achievement Awards Recognition." May 7, 2024. 15. New York Times: "Are milk and eggs safe as bird flu spreads?" April 5, 2024. 16. Montague Reporter: "Endless Worries (heading) Regional Norovirus Outbreak: Should We Be Freaking Out?" Front Page. February 29, 2024. Perhaps most rewarding, PI Moore was selected as the sole rising star faculty for the UMass College of Natural Sciences by the Dean to speak at the UMass Chancellor's house for a high stakes donor VIP gathering in October 2024, where PI Moore spoke about a number of projects including this one.? What do you plan to do during the next reporting period to accomplish the goals?We plan to continue with publication of our second manuscript, as well as continue evaluating the performance of the sensors in other foods. We also aim to try to generate these sensors against aflatoxin for evaluation of sensitivity and ability to detect mycotoxins in foods.

Impacts
What was accomplished under these goals? We expanded upon understanding related to Objective 1 and 3 in the past year, especially attempting to nderstand the influences of a.) the strain of the target peptide used to create the nanoMIPs, as well as the influence on the degree of selectivity of the generated nanoMIPs to a broader array of related viral targets. Additionally, we were able to demonstrate that the developed nanoMIP technology was able to perform in food samples relevant to norovirus transimission (leafy greens). Despite an inherenet delay due to the co-PI moving to a new institution, we were able to wrap up our second manuscript. Norovirus is the leading cause of viral gastroenteritis worldwide, contributing to widespread disease and financial burdens. However, current testing methods are unsuitable for on-site analysis, as they typically use biological receptors, require specialized reagents, and skilled technicians. Proactive on-site testing of high-risk food samples is essential to prevent outbreaks. We have developed a thermal sensor capable of selectively detecting two recurrent norovirus genotypes, GI.1 and GII.4, within a model food matrix. The sensor utilizes epitope-imprinted polymer nanoparticles (nanoMIPs) designed from a 10-amino-acid sequence derived from the conserved P1 region of the GI.1 viral capsid (SEQAPTVGEA), with the generated particles determined to have a hydrodynamic diameter (Dh) of 178 nm. This was a different target than the first set of nanoMIPs that were generated in our first paper from previous years. The nanoMIPs demonstrated favorable sensitivity to norovirus GI.1 and GII.4 virus-like particles in buffer solutions, achieving detection limits of 1.53 and 2.28 pg/mL (0.87 × 105 and 1.30 × 105 particles/mL), respectively. This limit is comparable to many portable immunoassay-based detection schemes. The fact the sensor was able to detect such a broad range shows great promise for the use of nanoMIPs as recognition ligands for noroviruses, as this can be a particular challenge for traditional ligands like monoclonal antibodies. The selectivity of the nanoMIPs was evaluated against a panel of similar viruses, including murine norovirus, Tulane virus, and bacteriophage MS2, each of which showed a reduced signal. The thermal binding responses for murine norovirus, Tulane virus, and bacteriophage MS2 showed a statistically significant difference when compared to the GI.1 VLPs, reflecting the increased genetic divergence of these viruses. Murine Norovirus, while still belonging to the Norovirus genus, showed a reduction in ΔRth compared to GI.1 and GII.4. The genetic variations among viruses can be quantitatively assessed by analyzing differences in the amino acid sequences of their capsid proteins. Norovirus GII.4 and murine norovirus showed 44% and 40% amino acid similarity to GI.1, respectively. The selected epitope region exhibits similarities between GI.1 and GII.4, whereas the reduced response of murine norovirus may be due to much of the sequence similarity being located in the shell domain or non-binding regions, leaving key differences in the binding region critical for interaction with the sensor. Tulane virus capsid protein was calculated to have a 29% similarity which reflects the reduced response of the sensor. Bacteriophage MS2, the most genetically distant virus in the panel, displayed no significant similarity and produced the smallest change in thermal resistance, indicating selectivity of the nanoMIPs for members of the Caliciviridae family and reduced/little response to other viruses that have similar capsid symmetry and structural properties. Interestingly, the fact that some cross-reactivity was observed with murine norovirus and Tulane virus suggests that nanoMIPs may be well suited as ligands for broad detection of diverse pathogens in general. Notably, the sensor achieved rapid detection in spinach rinsate samples (30 min) while maintaining comparable detection limits (2.19 pg/mL and 2.69 pg/mL). This is promising in particular, as it suggests that nanoMIP sensors may be less prone to inhibition by food matrices, which is a major advantage in detecting viruses in foods in a portable manner, as it means that cruder and faster sample processing prior to detection may be possible. The combination of fast detection time, broad strain recognition, and straightforward sample preparation makes the nanoMIP thermal sensor a promising tool for on-site testing in food safety and public health settings.

Publications

  • Type: Peer Reviewed Journal Articles Status: Under Review Year Published: 2025 Citation: Dann A, Singla P, McClements J, Kim M, Stoufer S, Crapnell R, Banks CE, Blanford CF, Moore MD* and Peeters M*. 2024. Epitope-Imprinted Polymers for Rapid Norovirus Detection in Food Samples. Sensors & Actuators B: Chemical. (Under Review).
  • Type: Conference Papers and Presentations Status: Published Year Published: 2024 Citation: Dann A, Singla P, Stoufer S, Kim M, Sullivan M, Turner N, Geoghegan M, Seyedin S, Moore MD, McClements J, and Peeters M. Molecularly imprinted nanoparticles for the detection of norovirus. Society for Molecular Imprinting Annual Conference 2024. Verona, Italy. 6/20/2024.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2024 Citation: Dann A, Singla P, Stoufer S, Kim M, Sullivan M, Turner N, Geoghegan M, Seyedin S, McClements J, Peeters M, and Moore MD. Development and evaluation of low-cost, easily deployable molecularly imprinted polymer nanoparticles for agricultural viruses and toxins of concern. Nanoscale Science and Engineering for Agriculture and Food Systems Gordon Research Conference. Manchester, NH, USA. 6/25/2024.
  • Type: Peer Reviewed Journal Articles Status: Published Year Published: 2024 Citation: Kaur S, Singla P, Dann A, McClements J, Sullivan M, Kim M, Stoufer S, Dawson J, Crapnell R, Banks C, Turner N, Moore MD, Kaur I, Peeters M. 2024. Sensitive Electrochemical and Thermal Detection of Human Noroviruses Using Molecularly Imprinted Polymer Nanoparticles Generated against a Viral Target. ACS Applied Materials & Interfaces 16(38):5139751410.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2024 Citation: Moore MD. Advances in the Detection and Control of Foodborne Viruses. Future of Food Symposium. McGill University and the Consortium de Recherche et innovations en Bioproc�d�s industriels au Qu�bec (CRiBiQ). Montreal, Quebec, Canada. 5/16/2024.
  • Type: Other Status: Published Year Published: 2024 Citation: Moore MD. Moore Lab: Applied and Environmental Virology. CNS Rising Stars in Research Innovation  informal meeting with VIP UMass CNS alumni at the invitation of UMass Chancellor Reyes and CNS Dean Fox to discuss Moore lab research. Amherst, MA, USA. 11/16/2024.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2024 Citation: Moore MD. Moore Lab: Applied and Environmental Virology. UMass Food Science Advisory Board Meeting. Amherst, MA, USA. 4/19/2024.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2024 Citation: Peeters M. "Peeters Research Group." University of Sheffield, United Kingdom. 7 Feb. 2024.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2024 Citation: Peeters M. "Peeters Research Group." KU Leuven, Belgium. 29 Feb. 2024.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2024 Citation: Peeters M. "Polymers in an age of data." High Polymer Research Group 2024. 30 April 2024. *co-PI Peeters had to be invited for this conference in the first place and there is only one of about 6 academic presentations there, the rest are by industry partners. It has a format comparable to a Gordon conference and a major honor to be invited.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2024 Citation: Singh P, Peeters M. "Biomimetic engineering: innovations inspired by nature." MACRO2024: 50th World Polymer Congress. Warwick University, Coventry, England, United Kingdom. 3 July 2024.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2024 Citation: Moore MD. Moore Lab: Applied and Environmental Virology. Department of Defense Army Combat Feeding Division Soldier Center. Natick, MA, USA. 2/27/2024.


Progress 02/01/23 to 01/31/24

Outputs
Target Audience:The target audience reached by the work in this reporting period consists primarily of fellow academics and researchers in the fields of detection/analytical biochemistry, microbiology, materials and bioengineering, and food safety. Secondary audiences reached in the different presentations/communications/publications involves members of the food/food safety industry, government, and academia. In particular, our data related to the potential of this technology to rapidly detect viruses as well as more fundamental developments in MIP technology to detect antibiotics in foods would be of interest to the broader infectious disease, polymer science, and food safety communities as a whole--including those who are focused on pathogen/contaminant control from a clinical perspective. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?This was a very successful year for professional training activities. We conducted a mutual exchange to visit each others' universities during the spring/summer of 2023, where PI Moore and two members of the Moore lab visited PI Peeters' lab in Newcastle, UK, and learned about molecularly imprinted polymers, 3D printing, and construction of microfluidic electrochemical sensors. Dr. Moore also gave a seminar for the School of Engineering at Newcastle University, and met a number of potential future collaborators. This trip to Newcastle, was also supplemented with the data from this project being presented at the International Association for Food Protection (IAFP) European Symposium, just a few hours north in Aberdeen, Scotland. The technical talk that coincided with this project was selected as a finalist for the technical talk competition, and one of the students who was working on this grant, Sloane Stoufer, presented the work in the competition. Another student who was partially supported by this grant as well as another NIFA grant, also attended the conference and was also a finalist for the technical talk competition, which she ultimately was selected as the winner. Similarly, PI Peeters, Postdoctoral Research Fellow McClements, and a graduate fellow (Amy Dann) also came to the Moore lab to visit UMass. They learned about a number of techniques related to applied microbiology and virology. Dr. Peeters also gave a very well-attended symposium and met with a number of different potential faculty collaborators at UMass. In addition to meeting many new connections at each respective university, as well as learning new scientific techniques and principles related to the project, the members of the project also were enabled to attend and present their research at a number of conferences in their home countries and abroad. This resulted in over 10 presentations at local, national, and international levels for the personnel related to the project with a variety of audiences. One student associated with the project, Minji Kim, also successfully defended her PhD. Further, PI Moore is heavily involved in the World Society for Virology (Treasurer, Co-Director, and Organizer for meetings) and attended on separate travel funds to present the poster to a different audience (virologists) compared to those who would normally attend IAFP. Dr. Moore served as one of the organizers of the WSV 2023 conference, which occurred in the summer of 2023 in Riga, Latvia. PI Moore gave an invited talk at the conference that presented results from this work, which received a good amount of interest from attendees, especially given the stability and potential for the technology in lower resource settings. Additionally, both PIs Moore and Peeters have aided and supported applications for different awards and fellowships for students and postdoc funded by the project. This has resulted in the students associated with this project winning multiple awards, some of which include: an IAFP 2023 Student Travel Award, oSTEM conference student travel award, UMass Hultin Competition (2nd Place; senior PhD student technical talk competition), and one of the students, Sloane Stoufer, applying for and receiving a USDA Predoctoral Fellowship that will cover the rest of her PhD for a different project related to detection. How have the results been disseminated to communities of interest?As mentioned above, the results and accomplishments have all been presented through numerous products of traditional means, like a publication, textbook chapter, posters, technical talks, invited talks, and also through numerous features in the popular press, a number of which are listed below: UMass News: "Food science doctoral candidates awarded USDA fellowships for food safety research." August 17, 2023. UMass CNS Press Release: "UMass PhD Candidates Awarded USDA Fellowships to Improve Food Safety." August, 2023. UMass News Release: "Matthew Moore elected to a leadership role for the World Society of Virology," April 7, 2023. UMass News Release: "Technology aims to quickly detect foodborne contaminants outside the lab," June 6, 2022. What do you plan to do during the next reporting period to accomplish the goals?We plan to continue evaluating the potential of the P domain peptide sequences to serve as targets for generation of nanoMIPs, as the fact the generated nanoMIPs show affinity for assembled viral capsid (VLPs), P dimers, and are broadly reactive with many Caliciviruses but selective in that they do not react with other similarly structured virusesis quite exciting. We plan to evaluate the potential for cocktails of peptides to enable generation of more broadly reactive nanoMIPs for sensing. Given the potential of the generated nanoMIPs, we plan to further determine the range of reactivity of the nanoMIPs, as well as how those generated against a different strain perform. We also aim for their integration into low-cost, portable, and easy-to-use devices for in-field testing. Further, we aim to repeat the nanoMIP synthesis process to target mycotoxins after continuing optimization with norovirus. Depending on progress of developing a self-contained device with the nanoMIPs integrated, the Moore lab will evaluate the nanoMIP sensors generated against viruses in more complex matrices (foods).

Impacts
What was accomplished under these goals? We expanded upon understanding related to Objective 1 and 3 in the past year, building upon accomplishments from objective 1 in year 1. Specifically, we finished up work related to our first paper for norovirus sensing, where we saw some very favorable results. Further, we executed a planned exchange of knowledge and leveraged the international trip to present and expand our scope. The exchange occurred in spring/summer 2023 between the labs, that also will coincided with the partner PIs delivering research seminars and meeting with other faculty at each others' respective institutions (UMass Amherst, May 31-June 6; Newcastle University, April 27-May 2). During the exchange each lab wastrained in respective disciplines (Peeters and group wastrained in microbiology, virology, and food safety techniques at UMass, while Moore and group will be trained in molecularly imprinted polymer nanoparticle formation, engineering, and microfluidic techniques while at Newcastle University). In addition to the training exchange and seminars, the date of the UMass visit to Newcastle was chosen to coincide with the International Association for Food Protection European Symposium being held May 3-5 in Aberdeen, Scotland, which is relatively close to Newcastle, UK (See Products). During this meeting, a supporting PhD student, Minji Kim, won the Technical Talk competition, and this work was selected as a finalist as well in that competition where Sloane Stoufer presented out promising results. We were able to demonstrate multiple formulations of nanMIPs generated against the 10mer peptide were capable of binding full, assembled virus-like particles in a broad enough manner to interact with genogroups GI and GII, while remaining selectivity. We also demonstrated the ability of the nanMIP sensor to work in a food matrix (produce wash), and drafted and completed a publication that was submitted from this work. We have since created new formulations and are more closely evaluating the degree to which the generated nanoMIPs are broadly/reactive/selective, finding affinity to other genera of the Caliciviridae family but not reacting with structurally related viruses outside the family. From this work we have drafted another publication we plan to submit in the next period.

Publications

  • Type: Book Chapters Status: Published Year Published: 2023 Citation: Moore MD, Anderson J, Bisha B, Brehm-Stecher B. 2024. Sample Preparation for Detection of Microbiological and Chemical Analytes. Textbook Chapter. Encyclopedia of Food Safety, 2nd Edition, Vol. 3: 285-294. Ed. Geoffrey Smithers.
  • Type: Other Journal Articles Status: Other Year Published: 2024 Citation: Dann A, McClements J, Kaur I, Stoufer S, Kim M, Moore MD, Peeters M. Thermal Detection of Norovirus Using Molecularly Imprinted Polymers: Assessing Selectivity with Surrogates and Detection in Food Samples. In Preparation.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2023 Citation: 2021-08570: PARTNERSHIP: Development and evaluation of low-cost, easily deployable molecularly imprinted polymer nanoparticles for agricultural viruses and toxins of concern. USDA NIFA Nanotechnology for Food and Agriculture Project Director Meeting. Knoxville, TN, USA. 8/10/2023.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2023 Citation: Developments in detection and control of highly transmissible viral pathogens. World Society for Virology 2023 Meeting. Riga, Latvia. 6/16/2023.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2023 Citation: Moore Lab: Applied and Environmental Virology. Invited Seminar, UMass Biomedical Engineering Graduate Society Seminar. Amherst, MA, USA. 3/8/2023.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2023 Citation: Dann A, Kaur S, Stoufer S, Peeters M, Moore MD. Molecularly Imprinted Nanoparticles for the Electrochemical Detection of Norovirus. USDA NIFA Nanotechnology for Food and Agriculture Project Director Meeting. Knoxville, TN, USA. August 2023. Poster.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2023 Citation: Jamieson O, McClements J, Kaiya G, Stoufer S, Moore MD, Bell J, Perez-Padilla V, Rurack K, and Peeters. The Devolvement of Polymer-Based Sensors for Detecting Antibiotics in Food. International Association for Food Protection Annual Meeting 2023. Toronto, Canada. July 2023. Poster.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2023 Citation: Kaur S, McClements J, Singla P, Dann A, Sullivan MV, Turner MW, Stoufer S, Kim M, Moore MD, Kaur I, Peeters M. Development and evaluation of low-cost, easily deployable molecularly imprinted polymer for norovirus detection. International Association for Food Protection European Symposium 2023. Aberdeen, Scotland. May 2023. *Sarbjeet Kaur selected as finalist for Student Research Competition; Sloane Stoufer provided the talk in place for Sarbjeet, as she could not physically attend the symposium
  • Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Kaur S, Singla P, McClements J, Moore MD, Kaur I, and Peeters M. Development and evaluation of low-cost, easily deployable molecularly imprinted polymer technologies for norovirus detection. Poster. 17th Pacific Polymer Conference. Brisbane, Australia. December 2022.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2023 Citation: Peeters M. "Peeters Research Group." University of Sheffield, United Kingdom. 7 Feb. 2024.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2023 Citation: Peeters M. "Peeters Research Group." KU Leuven, Belgium. 29 Feb. 2024.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2024 Citation: Peeters M. "Polymers in an age of data." High Polymer Research Group 2024. 30 April 2024. *co-PI Peeters had to be invited for this conference in the first place and there is only one of about 6 academic presentations there, the rest are by industry partners. It has a format comparable to a Gordon conference and a major honor to be invited.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2024 Citation: Singh P, Peeters M. "Biomimetic engineering: innovations inspired by nature." MACRO2024: 50th World Polymer Congress. Warwick University, Coventry, England, United Kingdom. 3 July 2024.
  • Type: Peer Reviewed Journal Articles Status: Submitted Year Published: 2023 Citation: Kaur S, Singla P, Dann A, McClements J, Sullivan M, Kim M, Stoufer S, Dawson J, Crapnell R, Banks C, Turner N, Moore MD#*, Kaur I, Peeters M. 2024. Sensitive Electrochemical and Thermal Detection of Human Noroviruses Using Molecularly Imprinted Polymer Nanoparticles Generated against a Viral Target. ACS Applied Materials & Interfaces 16(38):5139751410.
  • Type: Book Chapters Status: Published Year Published: 2023 Citation: Dann A, Kaur S, Stoufer S, Kim M, Kaur I, Moore MD#, Peeters M, McClements J. 2023. Imprinted Polymers for Detection of Chemical and Microbial Contaminants in Foods. Textbook Chapter. Encyclopedia of Food Safety, 2nd Edition, Vol. 3:303-314. Ed. Geoffrey Smithers.


Progress 02/01/22 to 01/31/23

Outputs
Target Audience:The target audience reached by the work in this reporting period consists primarily of fellow academics and researchers in the fields of detection/analytical biochemistry, microbiology, materials and bioengineering, and food safety. Secondary audiences reached in the different presentations/communications/publications involves members of the food/food safety industry, government, and academia. In particular, our data related to the potential of this technology to rapidly detect viruses as well as more fundamental developments in MIP technology to detect antibiotics in foods would be of interest to the broader infectious disease, polymer science, and food safety communities as a whole--including those who are focused on pathogen/contaminant control from a clinical perspective. Changes/Problems:The only problems encountered so far were related to scheduling, as originally the exchange was planned for reporting period 1; however, based on availability and timing, as well as research progress, the exchange was pushed back to the spring of 2023 and is anticipated to take place in April and May 2023. What opportunities for training and professional development has the project provided?We have scheduled a delayed exchange for spring 2023 between the labs, that also will coincide with the partner PIs delivering research seminars and meeting with other faculty at each others' respective institutions (UMass Amherst, May 31-June 6; Newcastle University, April 27-May 2). During the exchange each lab will be trained in respective disciplines (Peeters and group will be trained in microbiology, virology, and food safety techniques at UMass, while Moore and group will be trained in molecularly imprinted polymer nanoparticle formation, engineering, and microfluidic techniques while at Newcastle University). In addition to the training exchange and seminars, the date of the UMass visit to Newcastle was chosen to coincide with the International Association for Food Protection European Symposium being held May 3-5 in Aberdeen, Scotland, which relatively close to Newcastle, UK (See Products). Additionally, a number of graduate students and postdocs will get/have had opportunities to present this work in conferences held in North America (IAFP Annual Meeting in Toronto), Australia (Technical Talk,), Scotland (IAFP European Symposium), and Latvia (World Society for Virology). Based on timing as well as the availability of additional discretionary funds of the PI for travel, we have been able to leverage attendance at additional conferences this year. For example, PI Moore is heavily involved in the World Society for Virology (Treasurer, Co-Director, and Organizer for meeting) and will be able to attend on separate travel funds to present the poster to a different audience (virologists) compared to those who would normally attend IAFP. Similarly, the Peeters group presented the project to a polymer science audience. Two graduate students from the Moore lab, as well as members of the Peeters lab, will get to attend the IAFP European Symposium in Aberdeen, Scotland, as a consequence of the location being close to Newcastle, UK, and timed with the UMass visit for the expertise exchange. Additionally, both PIs Moore and Peeters have aided and supported applications for different awards and fellowships for students and postdoc funded by the project. Including a Fellowship from the Royal Society of Chemistry for which the postdoc has made the final round (still awaiting decision), and a prestigious fellowship to one of the graduate students at UMass (UMass Food Science Manley Fellowship). How have the results been disseminated to communities of interest?As mentioned above, we either have abstracts submitted or have presented this work to numerous scientific and academic communities internationally. We also have a textbook chapter under review for the food safety community, and a peer-reviewed manuscript under minor revision related to the same nanoMIP development for a food contaminant (Products). What do you plan to do during the next reporting period to accomplish the goals?We plan to continue evaluating the potential of the P domain peptide sequences to serve as targets for generation of nanoMIPs, as the fact the generated nanoMIPs show affinity for assembled viral capsid (VLPs) is quite exciting. We plan to evaluate the potential for cocktails of peptides to enable generation of more broadly reactive nanoMIPs for sensing. Additionally, we will covalently functionalize the norovirus nanoMIPs to low-cost and highly reproducible SPEs using previously established methods. These functionalized SPEs can then be utilized for electrochemical detection of norovirus with the potential for integration into low-cost, portable, and easy-to-use devices for in-field testing. Further, we aim to repeat the nanoMIP synthesis process to target mycotoxins after continuing optimization with norovirus. Depending on progress and after training/exchange, the Moore lab will attempt to evaluate the nanoMIP sensors generated against viruses in more complex matrices (foods).

Impacts
What was accomplished under these goals? We have investigated and generated a number of different nanoMIP formulations against a norovirus epitope on the exposed outer portion of a norovirus epidemic genotype (GII.4) capsid protein (YQEAAPAQSDV) as the target for NanoMIP synthesis. This allowed for low-cost and safe synthesis as only a tiny virus fragment was required to develop the synthetic receptors; if successful, this could serve as a much cheaper and more efficient means of generating norovirus-specific ligands, as well as generate broadly reactive ligands against a cocktail of different strains. NanoMIP synthesis begins by immobilizing the target (norovirus epitope) to functionalized glass beads, which act as a solid-phase support during synthesis. The monomers are then allowed to self-assemble around the target before crosslinkers/initiators are added to lock the polymer structure in place. A low-temperature elution is initially performed with room-temperature water to remove any unreacted monomers or low-affinity nanoMIPs from the solution. This is followed by a high-temperature elution (70 °C), which separates the high-affinity nanoMIPs from the target. The collected high-affinity nanoMIPs contain cavities within them that are the correct size, shape, and functionality to selectively rebind with the target upon exposure. Essentially, they mimic the lock-and-key mechanism observed in biology. To develop high-performance nanoMIPs, it is vital to optimize monomer selection. This can be performed experimentally or by using computational techniques. Wherein, we developed innovative nanoMIPs using electroactive monomers [N-Isopropylacrylamide (NIPAM), N-(Tert-Butyl)Acrylamide (TBAM), Ferrocenylmethyl methacrylate (FMMA), N-(3-Aminopropyl)methacrylamide hydrochloride (NAMPA), Acrylic acid (AAc), and Dopamine Methacrylamide (DPMA)], along with a cross-linker [N, N'-methylenebisacrylamide (BIS)]. This means that they can be utilized for electrochemical detection which is optimal for portable, low-cost, and rapid in-field testing. Three batches of nanoMIPs were developed (two different electroactive and one non-electroactive). As these nanoMIPs are novel, the protocol development and optimization accounted for a considerable time period. NIPAM (mg) TBAM (mg) FMMA (mg) DPMA (mg) NAMPA (mg) AAc (µL) BIS (mg) TEMED (µL) APS (mg) Batch 1 (Standard Batch) 20 17 - - 4 1.1 1.5 15 24 Batch 2 (Ferrocene Batch) 20 17 20 - 4 1.1 1.5 15 24 Batch 3 (Dopamine Batch) 20 17 - 20 4 1.1 1.5 15 24 As confirmed by SEM, the nanoMIPs showed spherical morphology. Furthermore, DLS showed relatively homogenous nanoMIPs with average sizes of 90, 100, and 110 nm for the standard, ferrocene, and dopamine nanoMIPs, respectively. This characterization confirms the nanoMIP synthesis protocol was effective and that the nanoMIPs possess the necessary morphology for favorable sensing performance. SPR was performed on the three types of prepared nanoMIPs. The results demonstrate that all nanoMIP types showed high binding affinities for the norovirus epitope, P-domain, and VLPs. Furthermore, measurements were performed using a similar epitope (non-target) and binding affinity was two orders of magnitude larger. Consequently, this demonstrates that the prepared nanoMIPs can selectively bind to a range of norovirus targets (different sizes) with high affinity. Sample Epitope KD (M) Selectivity KD (M) P-domain KD (M) VLPs KD (M) Standard Batch 3.28 × 10-7 - 6.65 × 10-7 5.12 × 10-7 Ferrocene Batch 7.50 × 10-7 2.67 × 10-5 5.75 × 10-7 7.95 × 10-7 Dopamine Batch 1.92 × 10-6 3.51 × 10-5 1.37 × 10-6 1.76 × 10-6 Thermal detection (heat transfer method) was used to confirm the sensing capabilities of the standard nanoMIPs (non-electroactive). The method shows receptor-target interactions through increases in thermal resistance (Rth) at the functionalized-electrode surface. More target binding = greater thermal resistance. The results above clearly show there is no statistically significant difference in Rth during multiple PBS additions. However, when a VLP-spiked PBS solution (100 ng/mL) is added, the thermal resistance increases significantly, whereby it is many times greater than the standard deviation of the baseline signal (3x and 6x baseline SD on graph). This demonstrates that nanoMIPs can detect the VLPs using thermal detection methods.

Publications

  • Type: Journal Articles Status: Under Review Year Published: 2023 Citation: Singla, P.; Kaur, S.; Jamieson, O.; Dann, A.; Garg, S.; Mahon, C.; Crapnell, R.D.; Banks, C.E.; Kaur, I.; Peeters, M. Electrochemical�and�Thermal Detection of Allergenic Substance Lyzosyme with Molecularly Imprinted Nanoparticles. Analytical and Bioanalytical Chemistry, 2023, In revision  minor revisions.
  • Type: Book Chapters Status: Awaiting Publication Year Published: 2023 Citation: Dann, A.; Kaur, S.; Singla, P.; Stoufer, S.; Kim, M.; Kaur, I.; Moore, M.D.; Peeters, M.; McClements, J. Molecularly� Imprinted Polymers for Detection of Chemical and Microbial Contaminants in Foods. Encyclopedia of Food Safety, 2nd Edition, 2023, Under Editorial Review.
  • Type: Conference Papers and Presentations Status: Under Review Year Published: 2023 Citation: Jamieson, O.; McClements, J.; Kaiya, G.E.; Stoufer, S.; Moore, M.D.; Bell, J.; P�rez-Padilla, V.; Rurack, K.; Peeters, M. The Devolvement of Polymer-based Sensors for Detecting Antibiotics in Food. International Association for Food Protection Annual Meeting, 07/16/23-07/19/23, Toronto, Ontario, Canada. Oral abstract submitted.
  • Type: Conference Papers and Presentations Status: Under Review Year Published: 2023 Citation: Kaur, S.; McClements, J.; Singla, P.; Dann, A.; Minji, K.; Stoufer, S.; Moore, M.D.; Kaur, I.; Peeters, M. Development and Evaluation of Low-Cost, Easily Deployable Molecularly Imprinted Polymers for Norovirus Detection. International Association for Food Protection European Symposium on Food Safety, 05/03/23-05/05/23, Aberdeen, Scotland, United Kingdom. Oral abstract submitted.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Kaur, S.; Singla, P.; McClements, J.; Minji, K.; Stoufer, S.; Moore, M.D.; Kaur, I.; Peeters, M. Development of Molecularly Imprinted Polymer (MIP) Technologies for Norovirus Detection. The 17th Pacific Polymer Conference, 11/12/22-14/12/22, Brisbane, Queensland, Australia. Oral presentation delivered.
  • Type: Conference Papers and Presentations Status: Under Review Year Published: 2023 Citation: Kaur, S.; Singla, P.; McClements, J.; Minji, K.; Stoufer, S.; Moore, M.D.; Kaur, I.; Peeters, M. Application of Molecularly Imprinted Polymer Nanoparticles for Viral Pathogen Detection. World Society for Virology Meeting 2023, 15/6/2023-17/6/2023, Riga, Latvia. Poster, Abstract Under Review.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Moore MD. Developments in Detection and Control of Viral Pathogens. Invited Talk, Advanced Strategies to Control Microorganisms in Seafood Session. Korean Society of Food Science and Technology Annual Meeting 2022. Busan, South Korea. 7/6/2022.