More than magnetic isolation: Dynabeads as strong Raman reporters towards simultaneous capture and identification of targets

Jongwan Lee; Marissa McDonald; Nikiwe Mhlanga; Jeon Woong Kang; Rohit Karnik; Loza F Tadesse

More than magnetic isolation: Dynabeads as strong Raman reporters towards simultaneous capture and identification of targets

ArXiv [Preprint]. 2023 Jul 22:arXiv:2305.07199v2.

Authors

Jongwan Lee¹, Marissa McDonald², Nikiwe Mhlanga¹, Jeon Woong Kang³, Rohit Karnik¹, Loza F Tadesse^{1

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Affiliations

¹ Department of Mechanical Engineering, MIT, Cambridge, MA, 02139, United States.
² Department of Health Sciences & Technology, MIT, Cambridge, MA, 02139, United States.
³ Laser Biomedical Research Center, G. R. Harrison Spectroscopy Laboratory, MIT, Cambridge, MA, 02139, United States.
⁴ Ragon Institute of Massachusetts General Hospital, MIT and Harvard, Cambridge, MA, 02139, United States.
⁵ Jameel Clinic for AI & Healthcare, MIT, Cambridge, MA, 02139, United States.

PMID: 37214136
PMCID: PMC10197730

Abstract

Dynabeads are superparamagnetic particles used for 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 makes 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 to 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 bi-layer, they have 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.

Keywords: Raman reporters; Salmonella enterica; foodborne illness; immunomagnetic Dynabeads.

Publication types

Preprint