More than magnetic isolation: Dynabeads as strong Raman reporters toward simultaneous capture and identification of targets
Name
J Raman Spectroscopy - 2023 - Lee - More than magnetic isolation Dynabeads as strong Raman reporters toward simultaneous.pdf
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6.94 MB
Format
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Checksum (MD5)
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Author(s)
Lee, Jongwan
McDonald, Marissa
Mhlanga, Nikiwe
Kang, Jeon Woong
Karnik, Rohit
Tadesse, Loza F.
Date Issued
August 29, 2023
Journal
Journal of Raman Spectroscopy
Publisher
Wiley
Citation
J. Lee, M. McDonald, N. Mhlanga, J. W. Kang, R. Karnik, L. F. Tadesse, J Raman Spectrosc 2023, 54(9), 905.
Version
Final published version
Abstract
Dynabeads are superparamagnetic particles used for the immunomagnetic purification of cells and biomolecules. Post‐capture, however, target identification relies on tedious culturing, fluorescence staining, and/or target amplification. Raman spectroscopy presents a rapid detection alternative, but current implementations target cells themselves with weak Raman signals. We present antibody‐coated Dynabeads as strong Raman reporter labels whose effect can be considered a Raman parallel of immunofluorescent probes. Recent developments in techniques for separating target‐bound Dynabeads from unbound Dynabeads make such an implementation feasible with high specificity. We deploy Dynabeads anti‐Salmonella to bind and identify Salmonella enterica, a major foodborne pathogen. Dynabeads present major peaks around 1000 and 1600 cm−1 from aliphatic and aromatic C‐C stretching of the polystyrene coating and near 1350 cm−1 from the ɣ‐Fe2O3 and Fe3O4 core, confirmed with electron dispersive X‐ray (EDX) imaging. Minor to no contributions are made from the surface antibodies themselves as confirmed by Raman analysis of surface‐activated, antibody‐free beads. Dynabeads' Raman signature can be measured in dry and liquid samples even at single shot ~30 × 30 μm area imaging using 0.5 s, 7‐mW laser acquisition with single and clustered beads providing a 44‐ and 68‐fold larger Raman intensity compared with the signature from cells. Higher polystyrene and iron oxide content in clusters yields larger signal intensity, and conjugation to bacteria strengthens clustering as a bacterium can bind to more than one bead as observed via transmission electron microscopy (TEM). Our findings shed light on the intrinsic Raman reporter nature of Dynabeads. When combined with emerging techniques for the separation of target‐bound Dynabeads from unbound Dynabeads such as using centrifugation through a density media bilayer, they have the potential to demonstrate their dual function for target isolation and detection without tedious staining steps or unique plasmonic substrate engineering, advancing their applications in heterogeneous samples like food, water, and blood.
Subjects
Spectroscopy
General Materials Science
MIT Department
Massachusetts Institute of Technology. Department of Mechanical Engineering
Harvard University--MIT Division of Health Sciences and Technology
Massachusetts Institute of Technology. Spectroscopy Laboratory
Ragon Institute of MGH, MIT and Harvard
Terms of Use
Creative Commons Attribution
Persistent DSpace Link
DOI of Published Version
https://doi.org/10.1002/jrs.6584
