Use of magnetic nanotrap particles in capturing Yersinia pestis virulence factors, nucleic acids and bacteria
Abstract Background Many pathogens, including Yersinia pestis, cannot be consistently and reliably cultured from blood. New approaches are needed to facilitate the detection of proteins, nucleic acid and microorganisms in whole blood samples to improve downstream assay performance. Detection of biom...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2021-06-01
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| Series: | Journal of Nanobiotechnology |
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| Online Access: | https://doi.org/10.1186/s12951-021-00859-8 |
| _version_ | 1828119485092986880 |
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| author | Alexandra N. Ii Shih-Chao Lin Benjamin Lepene Weidong Zhou Kylene Kehn-Hall Monique L. van Hoek |
| author_facet | Alexandra N. Ii Shih-Chao Lin Benjamin Lepene Weidong Zhou Kylene Kehn-Hall Monique L. van Hoek |
| author_sort | Alexandra N. Ii |
| collection | DOAJ |
| description | Abstract Background Many pathogens, including Yersinia pestis, cannot be consistently and reliably cultured from blood. New approaches are needed to facilitate the detection of proteins, nucleic acid and microorganisms in whole blood samples to improve downstream assay performance. Detection of biomarkers in whole blood is difficult due to the presence of host proteins that obscure standard detection mechanisms. Nanotrap® particles are micron-sized hydrogel structures containing a dye molecule as the affinity bait and used to detect host biomarkers, viral nucleic acids and proteins as well as some bacterial markers. Nanotraps have been shown to bind and enrich a wide variety of biomarkers and viruses in clinically relevant matrices such as urine and plasma. Our objective was to characterize the binding ability of Nanotrap particle type CN3080 to Y. pestis bacteria, bacterial proteins and nucleic acids from whole human blood in order to potentially improve detection and diagnosis. Results CN3080 Nanotraps bind tightly to Yersinia bacteria, even after washing, and we were able to visualize the co-localized Nanotraps and bacteria by electron microscopy. These magnetic hydrogel Nanotraps were able to bind Yersinia DNA, supporting the utility of Nanotraps for enhancing nucleic acid-based detection methods. Nanotraps were capable of increasing Y. pestis nucleic acid yield by fourfold from whole human blood compared to standard nucleic acid extraction. Interestingly, we found CN3080 Nanotraps to have a high affinity for multiple components of the Yersinia type III secretion system (T3SS), including chaperone proteins, Yop effector proteins and virulence factor protein LcrV (V). Using Nanotraps as a rapid upstream sample-prep tool, we were able to detect LcrV in human blood by western blotting with minimal blood interference in contrast to direct western blotting of blood samples in which LcrV was obscured. We were able to computationally model the interaction of LcrV with the CN3080 Nanotrap dye and found that it had a low delta-G, suggesting high affinity. Importantly, Nanotraps were also able to enhance detection of secreted Yersinia proteins by mass spectrometry. Conclusion Upstream use of magnetic CN3080 Nanotrap particles may improve the downstream workflow though binding and enrichment of biomarkers and speed of processing. Utilization of Nanotrap particles can improve detection of Yersinia pestis proteins and nucleic acid from whole human blood and contribute to downstream assays and diagnostics including molecular methods such as sequencing and PCR and protein-based methods. Graphic Abstract |
| first_indexed | 2024-04-11T13:49:45Z |
| format | Article |
| id | doaj.art-0d8d339a0f22468684c9ddbdfc6ab610 |
| institution | Directory Open Access Journal |
| issn | 1477-3155 |
| language | English |
| last_indexed | 2024-04-11T13:49:45Z |
| publishDate | 2021-06-01 |
| publisher | BMC |
| record_format | Article |
| series | Journal of Nanobiotechnology |
| spelling | doaj.art-0d8d339a0f22468684c9ddbdfc6ab6102022-12-22T04:20:42ZengBMCJournal of Nanobiotechnology1477-31552021-06-0119111710.1186/s12951-021-00859-8Use of magnetic nanotrap particles in capturing Yersinia pestis virulence factors, nucleic acids and bacteriaAlexandra N. Ii0Shih-Chao Lin1Benjamin Lepene2Weidong Zhou3Kylene Kehn-Hall4Monique L. van Hoek5School of Systems Biology, George Mason UniversitySchool of Systems Biology, George Mason UniversityCeres NanosciencesCenter for Applied Proteomics and Personalized Medicine, George Mason UniversitySchool of Systems Biology, George Mason UniversitySchool of Systems Biology, George Mason UniversityAbstract Background Many pathogens, including Yersinia pestis, cannot be consistently and reliably cultured from blood. New approaches are needed to facilitate the detection of proteins, nucleic acid and microorganisms in whole blood samples to improve downstream assay performance. Detection of biomarkers in whole blood is difficult due to the presence of host proteins that obscure standard detection mechanisms. Nanotrap® particles are micron-sized hydrogel structures containing a dye molecule as the affinity bait and used to detect host biomarkers, viral nucleic acids and proteins as well as some bacterial markers. Nanotraps have been shown to bind and enrich a wide variety of biomarkers and viruses in clinically relevant matrices such as urine and plasma. Our objective was to characterize the binding ability of Nanotrap particle type CN3080 to Y. pestis bacteria, bacterial proteins and nucleic acids from whole human blood in order to potentially improve detection and diagnosis. Results CN3080 Nanotraps bind tightly to Yersinia bacteria, even after washing, and we were able to visualize the co-localized Nanotraps and bacteria by electron microscopy. These magnetic hydrogel Nanotraps were able to bind Yersinia DNA, supporting the utility of Nanotraps for enhancing nucleic acid-based detection methods. Nanotraps were capable of increasing Y. pestis nucleic acid yield by fourfold from whole human blood compared to standard nucleic acid extraction. Interestingly, we found CN3080 Nanotraps to have a high affinity for multiple components of the Yersinia type III secretion system (T3SS), including chaperone proteins, Yop effector proteins and virulence factor protein LcrV (V). Using Nanotraps as a rapid upstream sample-prep tool, we were able to detect LcrV in human blood by western blotting with minimal blood interference in contrast to direct western blotting of blood samples in which LcrV was obscured. We were able to computationally model the interaction of LcrV with the CN3080 Nanotrap dye and found that it had a low delta-G, suggesting high affinity. Importantly, Nanotraps were also able to enhance detection of secreted Yersinia proteins by mass spectrometry. Conclusion Upstream use of magnetic CN3080 Nanotrap particles may improve the downstream workflow though binding and enrichment of biomarkers and speed of processing. Utilization of Nanotrap particles can improve detection of Yersinia pestis proteins and nucleic acid from whole human blood and contribute to downstream assays and diagnostics including molecular methods such as sequencing and PCR and protein-based methods. Graphic Abstracthttps://doi.org/10.1186/s12951-021-00859-8Yersinia pestisDNANucleic acidProteinProteomicsSample preparation |
| spellingShingle | Alexandra N. Ii Shih-Chao Lin Benjamin Lepene Weidong Zhou Kylene Kehn-Hall Monique L. van Hoek Use of magnetic nanotrap particles in capturing Yersinia pestis virulence factors, nucleic acids and bacteria Journal of Nanobiotechnology Yersinia pestis DNA Nucleic acid Protein Proteomics Sample preparation |
| title | Use of magnetic nanotrap particles in capturing Yersinia pestis virulence factors, nucleic acids and bacteria |
| title_full | Use of magnetic nanotrap particles in capturing Yersinia pestis virulence factors, nucleic acids and bacteria |
| title_fullStr | Use of magnetic nanotrap particles in capturing Yersinia pestis virulence factors, nucleic acids and bacteria |
| title_full_unstemmed | Use of magnetic nanotrap particles in capturing Yersinia pestis virulence factors, nucleic acids and bacteria |
| title_short | Use of magnetic nanotrap particles in capturing Yersinia pestis virulence factors, nucleic acids and bacteria |
| title_sort | use of magnetic nanotrap particles in capturing yersinia pestis virulence factors nucleic acids and bacteria |
| topic | Yersinia pestis DNA Nucleic acid Protein Proteomics Sample preparation |
| url | https://doi.org/10.1186/s12951-021-00859-8 |
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