Source: UNIVERSITY OF CENTRAL FLORIDA submitted to NRP
SIMPLE AND SENSITIVE IMMUNODETECTION OF ANIMAL-DERIVED ADULTERANTS IN FOODS USING CATALYTIC NANOPARTICLES
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
AFRI COMPETITIVE GRANT
Reporting Frequency
Annual
Accession No.
1022124
Grant No.
2020-67021-31257
Cumulative Award Amt.
$490,000.00
Proposal No.
2019-05845
Multistate No.
(N/A)
Project Start Date
Sep 1, 2020
Project End Date
Aug 31, 2025
Grant Year
2020
Program Code
[A1511]- Agriculture Systems and Technology: Nanotechnology for Agricultural and Food Systems
Recipient Organization
UNIVERSITY OF CENTRAL FLORIDA
12722 RESEARCH PARKWAY
ORLANDO,FL 32826
Performing Department
Chemistry
Non Technical Summary
Animal-derived adulterants (i.e., fraudulently added substance) in foods has been a critical issue as food fraud for years, which not only violates relevant food regulations but also relates to food quality and safety. In particular, animal-derived adulterants can cause foodborne illnesses such as individual allergies and infection of foodborne pathogens. Effective analytical techniques for adulterant detection are crucially important to monitor and prevent adulterated food products. Nevertheless, current analytical techniques often involve sophisticated instruments, complicate procedures, high cost, and/or skilled personnel, which inhibits the extensive implementation of adulterant detection in the food industry. This project aims to develop a simple paper-based test strip for the rapid and sensitive detection of animal-derived adulterants in foods. The test strip works in a similar way as the over-the-counter pregnancy test, ensuring great simplicity of the detection. Meanwhile, the test strip is highly sensitive owing to the use of carefully engineered nanoparticles that can generate strong detection signal. Such a test strip is expected to be widely used in the food industry because of its simplicity and cost-effectiveness. The success of this project will be of great benefit to fighting food fraud by providing a type of simple, affordable, and sensitive strip test for adulterant detection.
Animal Health Component
(N/A)
Research Effort Categories
Basic
100%
Applied
(N/A)
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
7233260200030%
7113520202040%
7113320202030%
Knowledge Area
711 - Ensure Food Products Free of Harmful Chemicals, Including Residues from Agricultural and Other Sources; 723 - Hazards to Human Health and Safety;

Subject Of Investigation
3320 - Meat, beef cattle; 3260 - Poultry meat; 3520 - Meat, swine;

Field Of Science
2020 - Engineering; 2000 - Chemistry;
Goals / Objectives
The over-arching goal of this project is to develop a simple and sensitive lateral flow assay (or test strip) based on catalytically active nanoparticles for the detection of animal-derived adulterants in food products. This goal will be addressed through the following three objectives: 1. Engineering the catalytic nanoparticles made of noble metals that possess optimized enzyme-like activities. The enzyme-like activities enable the nanoparticles to generate sensitive detection signal by catalyzing chromogenic substrates. 2. Establishing the catalytic nanoparticles-based test strip, aiming a substantial improvement (ideally two orders of magnitude) in detection sensitivity relative to conventional gold nanoparticles-based test strip. 3. Applying the catalytic nanoparticles-based test strip to detection of animal-derived adulterants in food products.
Project Methods
Approaches and methods for accomplishing the objectives are outlined in the following. In objective 1, the catalytic nanoparticles will be synthesized using a seed-mediated method, where preformed gold nanoparticles as seeds will be coated with a second noble metal. As-prepared nanoparticles will be characterized with electron microscopy, from which the size, shape, and elemental composition of the particles will be revealed. Enzyme-like catalytic activity of the nanoparticles will be evaluated using the steady-state kinetic assay. In objective 2, the catalytic nanoparticles will be functionalized with antibodies that are specific to certain analytes to form nanoparticle-antibody conjugates. These conjugates will be used as labels to assemble test strip. The test strip will be used to detect porcine skeletal troponins (from meat) and hemoglobin (from blood) as model analytes in assay buffer, from which the sensitivity, reproducibility, and stability of the test strip will be determined. In objective 3, Various mammalian meat and blood spiked food samples will be detected using the catalytic nanoparticles-based test strip. Non-specific signal of the detection will be evaluated and eliminated by optimizing assay conditions. Detection performance of the test strip will be evaluated by comparing it with commercially available detection kits.

Progress 09/01/23 to 08/31/24

Outputs
Target Audience: Nothing Reported Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?This project supported two graduate students (one at UCF and the other at FSU). With the support of this grant, the students have been trained with research skills of nanotechnology, materials science, and food science, and gained hand-on research experiences. How have the results been disseminated to communities of interest?The results have been disseminated to the community through peer-reviewed publications. Publications of current reporting period: 1. Gao, W.; Sun, X.; Yishay, T.; Wei, Z.; Zhu, X.; Kim, M.; Xia, X. Iridium nanoparticles as highly effective peroxidase mimics: synthesis, characterization, and application in biosensing. ChemNanoMat, 2024, 10, e202300589. 2. Shao, S.; Wang, X.; Sorial, C.; Sun, X.; Xia, X. Sensitive colorimetric lateral flow assays enabled by platinum-group metal nanoparticles with peroxidase-like activities. Advanced Healthcare Materials, 2024, in press, DOI: 10.1002/adhm.202401677. What do you plan to do during the next reporting period to accomplish the goals?The next reporting period will be the last period for this project. The research team will finalize the proposed lateral flow assay system and apply it to detection of adulterants in foods; At least one research paper will be submitted/published to peer-reviewed journals; Graduate students who are involved in this project will present their research findings at local and/or national conferences.

Impacts
What was accomplished under these goals? Major accomplishments in the current reporting period (09/2023-08/2024) include: i) Nanoparticles (NPs) made of platinum-group metals (PGM) have been demonstrated to possess superior peroxidase-like catalytic activities compared to many other materials. In our recent study, we found that iridium (Ir) is the most active PGM among others. The findings were published in ACS Applied Nano Materials 2022, 5, 17622-17631, which was reported in our last reporting period (09/2022-08/2023). On the basis of these recent scientific findings, we have prepared a class of unique iridium nanoparticles (Ir NPs) with rough surfaces that possess an ultrahigh peroxidase-like catalytic efficiency with a catalytic constant (Kcat) at the regime of 1010 s-1. The Ir NPs were used as labels to develop a new type of colorimetric lateral flow assay (CLFA). The Ir NPs-based CLFA achieved a low limit of detection (LoD), which was more than 20 times lower than the LoD of conventional gold nanoparticles-based CLFA. The results were published in ChemNanoMat, 2024, 10, e202300589. ii) The project director (PD) Dr. Xia's group was invited to publish a Perspective article in journal Advanced Healthcare Materials, entitled "Sensitive colorimetric lateral flow assays enabled by platinum-group metal nanoparticles with peroxidase-like activities". This article introduces the design, synthesis, and characterization of PGM NPs with peroxidase-like activities and the application of PGM NPs in development of CLFA tests. Research findings from this NIFA-supported project were incorporated into this article. This perspective article is expected to be a valuable resource for researchers in the areas of nanotechnology, biomedicine, and food and agriculture research. iii) The Co-PD Dr. Rao's research group has accomplished the followings: One manuscript related to fish allergen detection has been submitted to the Journal of Agricultural and Food Chemistry, which is currently under review. Four poster presentations were delivered at professional meetings, including the IFT Annual Meeting and the FAFP Annual Education Conference. One of these presentations was selected for an oral competition, where it earned second place in the Food Safety and Quality Management Division at the 2024 IFT FIRST Annual Event & Expo.

Publications

  • Type: Peer Reviewed Journal Articles Status: Published Year Published: 2024 Citation: Shao, S.; Wang, X.; Sorial, C.; Sun, X.; Xia, X. Sensitive colorimetric lateral flow assays enabled by platinum-group metal nanoparticles with peroxidase-like activities. Advanced Healthcare Materials, 2024, in press, DOI: 10.1002/adhm.202401677
  • Type: Peer Reviewed Journal Articles Status: Published Year Published: 2024 Citation: Gao, W.; Sun, X.; Yishay, T.; Wei, Z.; Zhu, X.; Kim, M.; Xia, X. Iridium nanoparticles as highly effective peroxidase mimics: synthesis, characterization, and application in biosensing. ChemNanoMat, 2024, 10, e202300589


Progress 09/01/22 to 08/31/23

Outputs
Target Audience: Nothing Reported Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?This project supported two graduate students (one at UCF and the other at FSU). With the support of this grant, these students have been trained with research skills of nanotechnology, food science, materials science and catalysis, and gained hand-on research experiences. How have the results been disseminated to communities of interest?The results have been disseminated to the community through peer-reviewed publications. Biby, A.; Crawford, H.; Xia, X. Platinum-group metal nanoparticles as peroxidase mimics: implications for biosensing. ACS Applied Nano Materials, 2022, 5, 17622-17631. Wei, Z.; Luciano, K.; Xia, X. Catalytic gold-iridium nanoparticles as labels for sensitive colorimetric lateral flow assay. ACS Nano, 2022, 16, 21609-21617. Jiang, X.; Rao, Q. Immunodetection of finfish residues on food contact surfaces. Food Chemistry 2023, 426, 136502. Gao, W.; Eastwood, H.; Xia, X. Peroxidase mimics of platinum-group metals for in vitro diagnostics: opportunities and challenges. Journal of Materials Chemistry B, 2023, 11, 8404-8410. What do you plan to do during the next reporting period to accomplish the goals?The PD and co-PD will work together to optimize the lateral flow assay system and apply it to detection of adulterants in foods; At least one research paper will be submitted/published to prestigious peer-reviewed journals; Graduate students who are involved in this project will present their research findings at local and/or national conferences.

Impacts
What was accomplished under these goals? Major accomplishments incurrent reporting period (09/2022-08/2023) include: i) Among various materials, noble metal (especially platinum-group metals, PGM) nanoparticles (NPs) have been demonstrated to possess superior peroxidase-like catalytic activities. Nevertheless, there is a lack of a comprehensive study that compares the peroxidase-like activities of PGM nanoparticles and their performance in biosensing. In a recent study, we have systematically studied PGM NPs as peroxidase mimics, including Pd, Pt, Rh, and Ir NPs. The results indicated that Ir NP is the most active one with a catalytic constant as high as 6.27 × 105 s-1, followed by Pt, Rh, and Pd NPs. Accordingly, Ir NPs showed the best performance in enzyme-linked immunosorbent assay as a model biosensing platform. The results were published in ACS Applied Nano Materials 2022, 5, 17622-17631. ii) Based on the abovementioned findings, Ir was chosen to coat on pre-formed Au nanoparticles (NPs) to form Au@Ir core@shell NPs. This design is consistent with the plan of original proposal, where Au@M core@shell (M = Pt, Ir, Pd, or Rh) NPs were proposed to be used as labels for colorimetric lateral flow assay (CLFA). The catalytic activity, in terms of catalytic constant Kcat, of the Au@Ir NPs was as high as 1.07 × 107 s-1. The highly catalytically active Au@Ir NPs as labels ensures a strong color signal and thus a high sensitivity of CLFA. The Au-Ir NPs-based CLFA was successfully applied to the detection of two different cancer biomarkers with limits of detection at the low picogram per milliliter level, which is hundreds of times more sensitive than conventional AuNP-based CLFA. The results were published in ACS Nano 2022, 16, 21609. iii) One manuscript related to fish allergen detection published in Food Chemistry 2023, 399, 133966. The detection was based on a competitive enzyme-linked immunosorbent assay (cELISA). The cELISA was successfully applied for quantification of major finfish allergen, parvalbumin, from swab samples. This work contributes to allergen surveillance in the food industry.

Publications

  • Type: Journal Articles Status: Published Year Published: 2022 Citation: Biby, A.; Crawford, H.; Xia, X. Platinum-group metal nanoparticles as peroxidase mimics: implications for biosensing. ACS Applied Nano Materials, 2022, 5, 1762217631
  • Type: Journal Articles Status: Published Year Published: 2022 Citation: Wei, Z.; Luciano, K.; Xia, X. Catalytic gold-iridium nanoparticles as labels for sensitive colorimetric lateral flow assay. ACS Nano, 2022, 16, 2160921617.
  • Type: Journal Articles Status: Published Year Published: 2023 Citation: Jiang, X.; Rao, Q. Immunodetection of finfish residues on food contact surfaces. Food Chemistry 2023, 426, 136502
  • Type: Journal Articles Status: Published Year Published: 2023 Citation: Gao, W.; Eastwood, H.; Xia, X. Peroxidase mimics of platinum-group metals for in vitro diagnostics: opportunities and challenges. Journal of Materials Chemistry B, 2023, 11, 84048410.


Progress 09/01/21 to 08/31/22

Outputs
Target Audience: Nothing Reported Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?This project supported two graduate students (one at UCF and the other at FSU). With the support of this grant, these students have been trained with research skills of nanotechnology, food science, materials science and catalysis, and gained hand-on research experiences. How have the results been disseminated to communities of interest?The results have been disseminated to the community through peer-reviewed publications. Two papers have been published and three manuscripts are under revision or preparation: 1. Type: Journal Articles Status: Published, Year Submitted: 2022, Citation: Zhao, Y., Jiang, X., Tang, C., & Rao, Q. Composition, structural configuration, and antigenicity of Atlantic cod (Gadus morhua) tropomyosin. Food Chemistry, 2023, 399, 133966. 2. Type: Journal Articles Status: Published, Year Submitted: 2022, Citation: Luciano, K., Wang, X., Liu, Y.; Eyler, E.; Qin, Z. & Xia, X. Noble Metal Nanoparticles for Point-of-Care Testing: Recent Advancements and Social Impacts. Bioengineering, 2022, 9(11), 666; https://doi.org/10.3390/bioengineering9110666. 3. Type: Journal Articles Status: In revision, Year Submitted: 2022, Citation: Wei, Z., Luciano, K., & Xia, X. Catalytic Gold-Iridium Nanoparticles as Labels for Sensitive Colorimetric Lateral Flow Assay. ACS Nano. 4. Type: Journal Articles Status: In revision, Year Submitted: 2022, Citation: Biby, A., Crawford, H., & Xia, X. Platinum-Group Metal Nanoparticles as Peroxidase Mimics: Implications for Biosensing. ACS Applied Nano Materials. 5. Type: Journal Articles Status: Under preparation, Year Submitted: N/A, Citation: Zhao, Y., Jiang, X., Bai, C., Xia, X., & Rao, Q. Monoclonal antibody-based competitive enzyme-linked immunosorbent assay for the quantification of finfish tropomyosin. What do you plan to do during the next reporting period to accomplish the goals?The PD and co-PD will continue working together to assess and optimize the lateral flow assay system and apply it to detection of adulterants in foods; At least one research paper will be submitted/published to prestigious peer-reviewed journals; The graduate students involved in this project will present their research findings at local, regional, and/or national conferences.

Impacts
What was accomplished under these goals? Major accomplishments in this reporting period (2021/09/01 - 2022/08/31) include: i) Nanoparticles made of various noble metals (including Pt, Ir, Pd, and Rh) that possess peroxidase-like catalytic activities were synthesized and systematically compared. The results suggest Ir nanoparticle among other noble meal nanoparticles displayed the strongest catalytic activity and showed better performance in biosensing. The manuscript about this research had been submitted to ACS Applied Nano Materials. The manuscript is currently under revision; Based on this initial finding, Ir was chosen to coat on pre-formed Au nanoparticles to form Au@Ir core@shell nanoparticles. Note, in this project, Au@M core@shell (M = Pt, Ir, Pd, or Rh) nanoparticles were proposed as labels for lateral flow assay. The Au@Ir nanoparticles were successfully applied to lateral flow assay that achieved a high sensitivity for detection of cancer biomarkers. The findings were summarized in a manuscript that had been submitted to a high-impact journal ACS Nano. The manuscript is currently under revision. In the near future, we will apply this Au@Ir nanoparticle-based lateral flow assay for detection of adulterants in foods. ii) The PD has published an invited Review Article in journal Bioengineering, entitled "Noble Metal Nanoparticles for Point-of-Care Testing: Recent Advancements and Social Impacts". Basic concept of noble-metal nanoparticle design and synthesis, and perspectives on their applications in point-of-care testing (especially lateral flow assay) were discussed in the article. This review article is expected to be a valuable resource for peer researchers in fields of nanotechnology, sensing, and diagnostics. iii) The Co-PD has accomplished the followings: Two indirect/direct competitive enzyme-linked immunosorbent assays (ELISAs) against finfish tropomyosin were established and compared using two different substrates (i.e., colorimetric and chemiluminescent).

Publications

  • Type: Journal Articles Status: Published Year Published: 2022 Citation: Yaqi Zhao; Xingyi Jiang; Chunya Tang; Qinchun Rao. Composition, structural configuration, and antigenicity of Atlantic cod (Gadus morhua) tropomyosin. Food Chemistry, 2023, 399, 133966.
  • Type: Journal Articles Status: Published Year Published: 2022 Citation: Keven Luciano; Xiaochuan Wang; Yaning Liu; Gabriella Eyler; Zhenpeng Qin; Xiaohu Xia. Noble Metal Nanoparticles for Point-of-Care Testing: Recent Advancements and Social Impacts. Bioengineering, 2022, 9(11), 666.


Progress 09/01/20 to 08/31/21

Outputs
Target Audience: Nothing Reported Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?This project supported one graduate students at UCF. With the support of this grant, the student has been trained with research skills of nanoscience, materials science and catalysis, and gained hand-on research experiences. How have the results been disseminated to communities of interest?The results have been disseminated to the community through peer-reviewed publications. One paper has been published and one is under review: 1.Type: Journal Articles. Status: Published, Year Published: 2020. Citation: Wei, Z.; Xi, Z.; Vlasov, S.; Ayala, J.; Xia, X. Nanocrystals of Platinum-Group Metals as Peroxidase Mimics for in Vitro Diagnostics. Chemical Communications, 2020, 56, 14962-14975. 2.Type: Journal Articles. Status: Under review, Year Submitted: 2021. Citation: Jiang, X., Zhao, Y., Tang, C., Appelbaum, M., Rao, Q. Aquatic Food Animals in the U.S.: Status Quo and Challenges. Comprehensive Reviews in Food Science and Food Safety. What do you plan to do during the next reporting period to accomplish the goals?The PD and co-PD willfurther evaluate and optimize the Au@Ir NP system and apply the Au@Ir NPs to CLFA platform; At least one research paper will be submitted/published to prestigious peer-reviewed journals; The graduate students involved in this project will present their research findings at local, regional, or national conferences.

Impacts
What was accomplished under these goals? i) After having tried various metals (M) in the proposed Au@MnL NP system, we have successfully identified and built a unique type of NPs that possess ultrahigh peroxidase-like catalytic activities. Specifically, such NPs are Au@Ir core@shell NPs that were synthesized by coating preformed Au NPs (as seeds) with Ir in the presence of sodium citrate as a colloidal stabilizer. Morphology, composition, and structure of the Au@Ir NPs were characterized and confirmed with various analytical tools. Significantly, peroxidase-like catalytic activity of the Au@Ir NPs, in terms of catalytic constant (Kcat, which is defined as the maximum number of colored molecules produced per second per NP), was determined to be approximately 1.2 × 107 s-1. This Kcat has met our originally aimed value in the proposal (i.e., ≥ 107 s-1). This result is significant because Kcat of most already reported catalytic nanoparticles with peroxidase-like activities is confined to 104-105 s-1. This finding is a breakthrough in the niche field of peroxidase mimic development. Such a high catalytic efficiency of Au@Ir NPs ensures intense color signal generated from the NPs through catalysis. Therefore, the Au@Ir NPs are expected to be excellent labels for the proposed CLFA of adulterants in foods (i.e., objectives 2 and 3 of proposed research). ii) The PD has published an invited Feature Article in a high-impact journal Chemical Communications, entitled "Nanocrystals of Platinum-Group Metals as Peroxidase Mimics for in Vitro Diagnostics" (Chem. Commun. 2020, 56, 14962-14975). Concept of catalytic nanoparticle design/synthesis and perspectives on their applications that are relevant to this project were included in the article. This publication is a valuable resource for peer scientists in general fields of nanotechnology, materials science, sensing and imaging. iii) The Co-PD at FSU has accomplished the followings:Anti-finfish tropomyosin monoclonal antibody was purified and characterized. The catalytic ability of goat anti-mouse IgG secondary antibody-conjugated nanoparticles and their selectivity were characterized and compared with horseradish peroxidase (HRP) labeled goat anti-mouse IgG secondary antibody through indirect non-competitive enzyme-linked immunosorbent assay (ELISA).

Publications

  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Wei, Zhiyuan; Xi, Zheng; Vlasov, Sergey; Ayala, Jasmin; Xia, Xiaohu. Nanocrystals of Platinum-Group Metals as Peroxidase Mimics for in Vitro Diagnostics. Chemical Communications, 2020, 56, 14962-14975.