Abstract
Recent advancements in the profiling of proteomes at the single-cell level necessitate the development of quantitative and versatile platforms, particularly for analyzing rare cells like circulating tumor cells (CTCs). In this chapter, we present an integrated microfluidic chip that utilizes magnetic nanoparticles to capture single tumor cells with exceptional efficiency. This chip enables on-chip incubation and facilitates in situ analysis of cell-surface protein expression. By combining phage-based barcoding with next-generation sequencing technology, we successfully monitored changes in the expression of multiple surface markers induced by CTC adherence. This innovative platform holds significant potential for comprehensive screening of multiple surface antigens simultaneously in rare cells, offering single-cell resolution. Consequently, it will contribute valuable insights into biological heterogeneity and human disease.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Orkin SH, Zon LI (2008) Hematopoiesis: an evolving paradigm for stem cell biology. Cell 132(4):631–644
Kreso A, Dick John E (2014) Evolution of the cancer stem cell model. Cell Stem Cell 14(3):275–291
Plaks V, Koopman CD, Werb Z (2013) Circulating tumor cells. Science 341(6151):1186–1188
Labib M, Kelley SO (2020) Single-cell analysis targeting the proteome. Nat Rev Chem 4(3):143–158
Li X, Fan B, Cao S, Chen D, Zhao X, Men D, Yue W, Wang J, Chen J (2017) A microfluidic flow cytometer enabling absolute quantification of single-cell intracellular proteins. Lab Chip 17(18):3129–3137
Palii CG, Cheng Q, Gillespie MA, Shannon P, Mazurczyk M, Napolitani G, Price ND, Ranish JA, Morrissey E, Higgs DR, Brand M (2019) Single-cell proteomics reveal that quantitative changes in co-expressed lineage-specific transcription factors determine cell fate. Cell Stem Cell 24(5):812–820.e815
Forbes TP, Forry SP (2012) Microfluidic magnetophoretic separations of immunomagnetically labeled rare mammalian cells. Lab Chip 12(8):1471–1479
Kang JH, Krause S, Tobin H, Mammoto A, Kanapathipillai M, Ingber DE (2012) A combined micromagnetic-microfluidic device for rapid capture and culture of rare circulating tumor cells. Lab Chip 12(12):2175–2181
Armbrecht L, Müller RS, Nikoloff J, Dittrich PS (2019) Single-cell protein profiling in microchambers with barcoded beads. Microsyst Nanoeng 5(1):55
Poudineh M, Sargent EH, Pantel K, Kelley SO (2018) Profiling circulating tumour cells and other biomarkers of invasive cancers. Nat Biomed Eng 2(2):72–84
Seemann R, Brinkmann M, Pfohl T, Herminghaus S (2012) Droplet based microfluidics. Rep Prog Phys 75(1):016601
Wu Q, Liu J, Wang X, Feng L, Wu J, Zhu X, Wen W, Gong X (2020) Organ-on-a-chip: recent breakthroughs and future prospects. Biomed Eng Online 19(1):9
Nie J, Gao Q, Wang Y, Zeng J, Zhao H, Sun Y, Shen J, Ramezani H, Fu Z, Liu Z, Xiang M, Fu J, Zhao P, Chen W, He Y (2018) Vessel-on-a-chip with hydrogel-based microfluidics. Small 14(45):1802368
Carvalho MR, Lima D, Reis RL, Correlo VM, Oliveira JM (2015) Evaluating biomaterial- and microfluidic-based 3D tumor models. Trends Biotechnol 33(11):667–678
van Duinen V, Trietsch SJ, Joore J, Vulto P, Hankemeier T (2015) Microfluidic 3D cell culture: from tools to tissue models. Curr Opin Biotechnol 35:118–126
Zhang W, Kai K, Choi DS, Iwamoto T, Nguyen YH, Wong H, Landis MD, Ueno NT, Chang J, Qin L (2012) Microfluidics separation reveals the stem-cell–like deformability of tumor-initiating cells. Proc Natl Acad Sci 109(46):18707–18712
Chen K, Dopico P, Varillas J, Zhang J, George TJ, Fan ZH (2019) Integration of lateral filter arrays with immunoaffinity for circulating-tumor-cell isolation. Angew Chem Int Ed 58(23):7606–7610
Cohen L, Walt DR (2019) Highly sensitive and multiplexed protein measurements. Chem Rev 119(1):293–321
Ledsgaard L, Ljungars A, Rimbault C, Sørensen CV, Tulika T, Wade J, Wouters Y, McCafferty J, Laustsen AH (2022) Advances in antibody phage display technology. Drug Discov Today 27(8):2151–2169
Cung K, Slater RL, Cui Y, Jones SE, Ahmad H, Naik RR, McAlpine MC (2012) Rapid, multiplexed microfluidic phage display. Lab Chip 12(3):562–565
Ma Y, Chen K, Xia F, Atwal R, Wang H, Ahmed SU, Cardarelli L, Lui I, Duong B, Wang Z, Wells JA, Sidhu SS, Kelley SO (2021) Phage-based profiling of rare single cells using nanoparticle-directed capture. ACS Nano 15(12):19202–19210
Author information
Authors and Affiliations
Corresponding authors
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2024 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Wang, Y., Zhao, J., Jiang, Z., Ma, Y., Zhang, R. (2024). Single-Cell Proteomics by Barcoded Phage-Displayed Screening via an Integrated Microfluidic Chip. In: Peng, H., Liu, J., Chen, I.A. (eds) Phage Engineering and Analysis. Methods in Molecular Biology, vol 2793. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3798-2_7
Download citation
DOI: https://doi.org/10.1007/978-1-0716-3798-2_7
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-3797-5
Online ISBN: 978-1-0716-3798-2
eBook Packages: Springer Protocols