DEVELOPMENT OF MAGNETO-PLASMONIC NANOSENSORS FOR THE ULTRASENSITIVE DETECTION OF FOOD-BORNE PATHOGENS

• Sponsoring Institution: National Institute of Food and Agriculture
• Project Status: COMPLETE
• Funding Source: AFRI COMPETITIVE GRANT
• Reporting Frequency: Annual
• Accession No.: 1018979
• Grant No.: 2019-67022-29457
• Cumulative Award Amt.: $145,015.00
• Proposal No.: 2018-07295
• Project Start Date: Aug 1, 2019
• Project End Date: Jul 31, 2022
• Grant Year: 2019
• Program Code: [A1511]- Agriculture Systems and Technology: Nanotechnology for Agricultural and Food Systems

Recipient Organization
PITTSBURG STATE UNIVERSITY
1501 S. JOPLIN STREET
PITTSBURG,KS 66762

Performing Department
Chemistry

Non Technical Summary
E. coli O157:H7, one of the leading causes of foodborne illness often results from consuming contaminated food and water. Its ability to produce Shiga-like toxins often lead to complications such as hemorrhagic colitis and hemolytic anemia. Although the Food and Drug Administration (FDA) does its best to prevent the sale of contaminated foods, the current detection methods aren't always a viable option for faster food testing. This issue is amplified in developing countries who don't always have the means to screen foods before consumption. Therefore, it is essential for researchers to find a fast, simple, and reliable method of bacterial detection for the benefit of producers and consumers alike. The goal of this study is to develop a simple magnetic relaxation platform with plasmonic properties that is user-friendly, rapid, and cost-effective and at the same time can be used in field conditions for food-safety applications. This technology would bring more sensitive methods for the effective detection of E. coli O157:H7 and other bacterial contaminations in food.

Animal Health Component

50%

Research Effort Categories

Basic

50%

Applied

50%

Developmental

(N/A)

Classification

Knowledge Area (KA)	Subject of Investigation (SOI)	Field of Science (FOS)	Percent
402	1430	2000	100%

Knowledge Area
402 - Engineering Systems and Equipment;

Subject Of Investigation
1430 - Greens and leafy vegetables;

Field Of Science
2000 - Chemistry;

Keywords

e. coli o157:h7

magneto-plasmonic nanosensors

nanozyme

mri

spr

food safety.

Goals / Objectives
The major goal of this project is to develop new multimodal nanosensor for the effective detection ofE. coli O157:H7, one of the leading causes of foodborne illness often results from consuming contaminated food. To meet this goal, this proposal introduces novel magneto-plasmonic nanosensors (MPnS) with integrated magnetic relaxation (T2 MR), plasmonic (SPR), colorimetric and enzymatic properties. All these unique properties in a single nanosensor platform makes it attractive for food-based pathogen sensing applications. In the present work, seed funding is requested for the development of this nanotechnology for the specific detection of E. coli O157:H7. In the MPnS design, gold nanoparticles (GNPs) encapsulating iron oxide nanoparticles (IONPs) will be formulated in one-step. E. coli O157:H7 has been selected as a model food-borne pathogen since it is infectious at low concentrations. The peroxidase-like activity of GNPs and IONPs will be compared with that of MPnS for the evaluation of possible synergistic catalytic activity.

Project Methods
Magnetic relaxation, surface plasmon resonance, peroxidase assay, nanozyme, ELISA, UV/Vis spectroscopy, colorimetry, nanotechnology, dialysis, dynamic light scattering, bacterial culture.

Progress 08/01/21 to 07/31/22

Outputs
Target Audience:Individual: University graduate and undergraduate students with learning and research experiences. Researchers interested in food-borne pathogens and related pathogen detections. Group and outreach: Part of this work was presented in Capitol Graduate Research Submit and undergraduate research days. The representatives of KBOR and research community were present in this summit. 1. "Development of Magneto-Plasmonic Nanosensors for the Detection of E. coli O157:H7". Eniola Arogunyo, Nilam Panchal, Carissa Sutton, Vedant Jain, Tuhina Banerjee, Santimukul Santra.* Capitol Graduate Research Summit, Topeka, KS, March 29, 2022. Poster presentation. 2. "Plasmon-Enhanced Bimodal Nanozyme: An Enzyme-Free Signal Amplification Strategy for Ultrasensitive Detection of Pathogens". Caine Duran, Nilam Panchal, Vedant Jain, Tuhina Banerjee, Santimukul Santra.* Kansas Undergraduate Research Days, Topeka, KS, February 29, 2022. Poster presentation. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Training activities: Graduate and undergraduate students received knowledge on various nanosensor formulations and characterizations. They received training on handling DLS, UV/Vis, magnetic relaxometer instruments, data collections and interpretation. They aquired knowledge on bacterial culture, E. coli culture, performing pathogen detection assays, ELIZA and Nanozyme experiments. Professional development: Students presented this work in ACS Midwest, K-INBRE and Capitol research summit in SP 2022 conferences. How have the results been disseminated to communities of interest?The results were disseminated to high school students visiting the university. Through presentations made by students and the PI: 1. "Biomimetic Macromolecular Nanoplatforms: Novel Biochemical Approaches for Rapidly Emerging Pathogens". Santimukul Santra. Department of Chemistry and Biochemistry, Missouri State University, February 16, 2022. Invited oral presentation. 2. "Development of Magneto-Plasmonic Nanosensors for the Detection of E. coli O157:H7". Eniola Arogunyo, Nilam Panchal, Carissa Sutton, Vedant Jain, Tuhina Banerjee, Santimukul Santra.* Capitol Graduate Research Summit, Topeka, KS, March 29, 2022. 3. "Plasmon-Enhanced Bimodal Nanozyme: An Enzyme-Free Signal Amplification Strategy for Ultrasensitive Detection of Pathogens". Caine Duran, Nilam Panchal, Vedant Jain, Tuhina Banerjee, Santimukul Santra.* Kansas Undergraduate Research Days, Topeka, KS, February 29, 2022. What do you plan to do during the next reporting period to accomplish the goals?1. Implication of MPnS sensor technology on other food-borne pathogen detections. 2. Establish lateral flow technology using MPnS nanosensor platform for the point-of-care detection of food-borne pathogens.

Impacts
What was accomplished under these goals? Novel nanosensor platform is developed for the specific and sensitive detection of E. coli O157:H7. This platform technology is successfully integrated to lateral flow assay (LFA) technology for field-deployable applications. This is now published in Analytical Chemistry journal, 2022, a highly cited peer reviewed journal in the field. Currently, this technology is under further development and we are working to receive another support from NIFA.

Publications

• Type: Journal Articles Status: Published Year Published: 2022 Citation: N. Panchal, V. Jain, R. Elliot, Z. Flint, W. Paul, D. Caine, T. Banerjee, S. Santra. "Plasmon-Enhanced Bimodal Nanosensor: An Enzyme-Free Signal Amplification Strategy for Ultrasensitive Detection of Pathogens". Analytical Chemistry, 2022, DOI: 10.1021/acs.analchem.2c03215.

Progress 08/01/19 to 07/31/22

Outputs
Target Audience:Individual: University graduate and undergraduate students with learning and research experiences. Researchers interested in food-borne pathogens and related pathogen detections. Farmers and growers affected with vegetable E.coli contaminations. Group and outreach: Part of this work was presented in Capitol Graduate Research Submit and undergraduate research days. The representatives of KBOR and research community were present in this summit. 1. "Development of Magneto-Plasmonic Nanosensors for the Detection of E. coli O157:H7". Eniola Arogunyo, Nilam Panchal, Carissa Sutton, Vedant Jain, Tuhina Banerjee, Santimukul Santra.* Capitol Graduate Research Summit, Topeka, KS, March 29, 2022. Poster presentation. 2. "Plasmon-Enhanced Bimodal Nanozyme: An Enzyme-Free Signal Amplification Strategy for Ultrasensitive Detection of Pathogens". Caine Duran, Nilam Panchal, Vedant Jain, Tuhina Banerjee, Santimukul Santra.* Kansas Undergraduate Research Days, Topeka, KS, February 29, 2022. Poster presentation. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Training activities: Graduate and undergraduate students received knowledge on various nanosensor formulations and characterizations. They received training on handling DLS, UV/Vis, magnetic relaxometer instruments, data collections and interpretation. They aquired knowledge on bacterial culture, E. coli culture, performing pathogen detection assays, ELIZA and Nanozyme experiments. Professional development: Students presented this work in ACS Midwest, K-INBRE and Capitol research summit in 2022 conferences. How have the results been disseminated to communities of interest?The results were disseminated to high school students visiting the university. Through presentations made by students and the PI: 1. "Biomimetic Macromolecular Nanoplatforms: Novel Biochemical Approaches for Rapidly Emerging Pathogens". Santimukul Santra. Department of Chemistry and Biochemistry, Missouri State University, February 16, 2022. Invited oral presentation. 2. "Development of Magneto-Plasmonic Nanosensors for the Detection of E. coli O157:H7". Eniola Arogunyo, Nilam Panchal, Carissa Sutton, Vedant Jain, Tuhina Banerjee, Santimukul Santra.* Capitol Graduate Research Summit, Topeka, KS, March 29, 2022. 3. "Plasmon-Enhanced Bimodal Nanozyme: An Enzyme-Free Signal Amplification Strategy for Ultrasensitive Detection of Pathogens". Caine Duran, Nilam Panchal, Vedant Jain, Tuhina Banerjee, Santimukul Santra.* Kansas Undergraduate Research Days, Topeka, KS, February 29, 2022. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? Novel nanosensor platform is developed for the specific and sensitive detection of E. coli O157:H7. This platform technology is successfully integrated to lateral flow assay (LFA) technology for field-deployable applications. This is now published in Analytical Chemistry journal, 2022, a highly cited peer reviewed journal in the field. Currently, this technology is under further development and we are working to receive another support from NIFA.

Publications

• Type: Journal Articles Status: Published Year Published: 2022 Citation: N. Panchal, V. Jain, R. Elliot, Z. Flint, W. Paul, D. Caine, T. Banerjee, S. Santra. "Plasmon-Enhanced Bimodal Nanosensor: An Enzyme-Free Signal Amplification Strategy for Ultrasensitive Detection of Pathogens". Analytical Chemistry, 2022, DOI: 10.1021/acs.analchem.2c03215.

Progress 08/01/20 to 07/31/21

Outputs
Target Audience:Individual: Graduate student(s) worked in this project gained research experiences and experimental learning. Group and outreach: Part of this work was presented in Capitol Graduate Research Submit, Topeka, KS on Feb 18th, 2021, The representatives of KBOR and research community were present in this summit. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Training activities: Students received knowledge on various nanosensor formulations and characterizations. Bacterial culture and handling for performing detection assays, ELIZA and Nanozyme experiments. Professional development: Students presented this work in K-INBRE and Capitol research summit in SP 2021 conferences. How have the results been disseminated to communities of interest?The results were disseminated to high school students visiting the university. However, these outreach efforts were restricted by the Covid-19 situations. Part of this project was presented by the PI at the NIFA Gordon Conference in 2021. What do you plan to do during the next reporting period to accomplish the goals?Optimizing Lateral Flow Assay (LFA) technology for the on-site detection of E.coli O157:H7 in higher sensitivity. Two manuscripts under preparation and will be submitted in Spring 2022.

Impacts
What was accomplished under these goals? New nanosensor platform was formulated with higher peroxidase-mimetic activity. Sandwitch ELISA using MPnS, new nanosensor, was successfully employed for detecting E.coli O157:H7.

Publications

• Type: Journal Articles Status: Published Year Published: 2019 Citation: ACS Appl. Nano Mater. 2019, 2, 5587?5593.

Progress 08/01/19 to 07/31/20

Outputs
Target Audience:The University and the community, by teaching, instructions and workshops. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Training activities: Students received knowledge on various nanosensors formulations and characterizations. Bacterial culture and handling for performing detection assays. Professional development: Students presented ithis work in K-INBRE and Capitol research summit in SP 2020 conferences. How have the results been disseminated to communities of interest?Summer workshop to school students (Greenbush, KS and Pittsburg High) for the GNPs synthesis and handson color change demonstrations. What do you plan to do during the next reporting period to accomplish the goals?More experiments on E.coli detection and collection of detection efficiency and limit. Peroxidase-like activity of MPnS and comparison with that of IONPs and GNPs for future applications.

Impacts
What was accomplished under these goals? Iron oxide nanoparticles were synthesized and characterized, showed superparamagnetic properties, data collected using Bruker's magnetic relaxometer. Gold nanoparticles synthesized and outstanding SPR properties were observed using TECAN's plate reaader. In-Situ magneto-plasmonic nanosensors were designed and synthesized and characterized using both relaxomether and plate reader. Size was determined using TEM. We were successful detecting E.coli in PBS and in milk complex media with a strong bi-modal signal (T2 and SPR) and color change from MPnS nanosensor. The sensitivity was over 10 fold higher than GNPs alone. Pathogen detection specificity experiments were performed using other food-borne bacteria species.

Publications

• Type: Journal Articles Status: Published Year Published: 2019 Citation: ACS Appl. Nano Mater. 2019, 2, 5587?5593.