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https://hdl.handle.net/2440/137334
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Type: | Journal article |
Title: | Magnetic enrichment of immuno-specific extracellular vesicles for mass spectrometry using biofilm-derived iron oxide nanowires |
Author: | Pham, Q.N. Winter, M. Milanova, V. Young, C. Condina, M.R. Hoffmann, P. Pham, N.T.H. Tung, T.T. Losic, D. Thierry, B. |
Citation: | Nanoscale, 2022; 15(3):1236-1247 |
Publisher: | Royal Society of Chemistry (RSC) |
Issue Date: | 2022 |
ISSN: | 2040-3364 2040-3372 |
Statement of Responsibility: | Quang Nghia Pham, Marnie Winter, Valentina Milanova, Clifford Young, Mark R. Condina, Peter Hoffmann, Nguyen T. H. Pham, Tran Thanh Tung, Dusan Losic and Benjamin Thierry |
Abstract: | Immuno-specific enrichment of extracellular vesicles (EVs) can provide important information into cellular pathways underpinning various pathologies and for non-invasive diagnostics, including mass spectrometry-based analyses. Herein, we report an optimised protocol for immuno-magnetic enrichment of specific EV subtypes and their subsequent processing with liquid chromatography-tandem mass spectrometry (LC-MS/MS). Specifically, we conjugated placental alkaline phosphatase (PLAP) antibodies to magnetic iron oxide nanowires (NWs) derived from bacterial biofilms and demonstrated the utility of this approach by enriching placenta-specific EVs (containing PLAP) from cell culture media. We demonstrate efficient PLAP+ve EV enrichment for both NW-PLAP and Dynabeads™-PLAP, with high PLAP protein recovery (83.7 ± 8.9% and 83.2 ± 5.9%, respectively), high particle-to-protein ratio (7.5 ± 0.7 × 109 and 7.1 ± 1.2 × 109 , respectively), and low non-specific binding of non-target EVs (7 ± 3.2% and 5.4 ± 2.2%, respectively). Furthermore, our optimized EV enrichment and processing approach identified 2518 and 2545 protein groups with LC-MS/MS for NW-PLAP and Dynabead™-PLAP, respectively, with excellent reproducibility (Pearson correlation 0.986 and 0.988). These findings demonstrate that naturally occurring iron oxide NWs have comparable performance to current gold standard immune-magnetic beads. The optimized immuno-specific EV enrichment for LC-MS/MS method provides a low-cost and highly-scalable yet efficient, high-throughput approach for quality EV proteomic studies. |
Keywords: | Placenta Humans Proteins Chromatography, Liquid Reproducibility of Results Proteomics Pregnancy Female Tandem Mass Spectrometry Nanowires Extracellular Vesicles |
Rights: | © The Royal Society of Chemistry 2023. |
DOI: | 10.1039/d2nr05619d |
Grant ID: | http://purl.org/au-research/grants/nhmrc/1171821 http://purl.org/au-research/grants/nhmrc/GNT1197173 http://purl.org/au-research/grants/arc/IH150100003 |
Published version: | http://dx.doi.org/10.1039/d2nr05619d |
Appears in Collections: | Chemical Engineering publications |
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