Analysis of Proteins at the Single Cell Level

Research output: Chapter in Book/Report/Conference proceedingChapterAcademicpeer-review

Abstract

Proteins bring a consequent additional level of information in comparison with nucleic acids on a cell’s state as proteins are dynamically processed and chemically modified in the cell as a function of the cell life. Yet, the analysis of proteins is challenging as no amplification step is possible as is the case for nucleic acids, and another difficulty lies in the dynamic range of protein expression in a single sample (e.g. a single cell). While the same challenges are still found for microfluidic-based analysis of proteins, microsystems bring about enhanced analytical performance and novel analysis opportunities. This is illustrated here for two different strategies that can be adopted for protein analysis in a chip format. A first strategy consists of transposing the standard proteomic protocol in miniaturized analytical tools, and this provides a number of advantages and enhancement for the analysis: an overall improvement is expected when using smaller systems whose capacity matches better the size of the samples; sample manipulation is minimized when using LOC technology,and this goes together with a decrease in sample loss and contamination; enhanced analytical performance in terms of analysis time and detection sensitivity is ensured by micro- and nano-scale features; last, the use of microfabricated structures guarantees higher analysis reproducibility. In a second strategy, the analysis is actually performed at the single cell level. This strategy does not enable protein mapping anymore, but the investigation focuses on given proteins (a single protein of a small number thereof) which are specifically targeted. For that purpose, innovative microfluidic-based protocols have been developed, and we classify them in three categories of fully destructive, partially invasive and non invasive protocols. Ongoing developments in the area of nanotechnology would enable truly protein mapping at the single cell level, with the use of nanofabricated tools in a LOC platform.
Original languageUndefined
Title of host publicationUnravelling Single Cell Genomics: Micro and Nanotools
EditorsNathalie Bontoux, Luce Dauphinot, Marie-Claude Potier
Place of PublicationCambridge
PublisherRoyal Society of Chemistry
Pages209-242
Number of pages34
ISBN (Print)978-1-84755-911-1
DOIs
Publication statusPublished - 2010

Publication series

NameRSC Nanoscience & Nanotechnology
PublisherRoyal Society of Chemistry
Number15
Volume15
ISSN (Print)1757-7136

Keywords

  • METIS-276315
  • EWI-19430
  • IR-75792

Cite this

le Gac, S. (2010). Analysis of Proteins at the Single Cell Level. In N. Bontoux, L. Dauphinot, & M-C. Potier (Eds.), Unravelling Single Cell Genomics: Micro and Nanotools (pp. 209-242). (RSC Nanoscience & Nanotechnology; Vol. 15, No. 15). Cambridge: Royal Society of Chemistry. https://doi.org/10.1039/9781849732284-00209
le Gac, Severine. / Analysis of Proteins at the Single Cell Level. Unravelling Single Cell Genomics: Micro and Nanotools. editor / Nathalie Bontoux ; Luce Dauphinot ; Marie-Claude Potier. Cambridge : Royal Society of Chemistry, 2010. pp. 209-242 (RSC Nanoscience & Nanotechnology; 15).
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le Gac, S 2010, Analysis of Proteins at the Single Cell Level. in N Bontoux, L Dauphinot & M-C Potier (eds), Unravelling Single Cell Genomics: Micro and Nanotools. RSC Nanoscience & Nanotechnology, no. 15, vol. 15, Royal Society of Chemistry, Cambridge, pp. 209-242. https://doi.org/10.1039/9781849732284-00209

Analysis of Proteins at the Single Cell Level. / le Gac, Severine.

Unravelling Single Cell Genomics: Micro and Nanotools. ed. / Nathalie Bontoux; Luce Dauphinot; Marie-Claude Potier. Cambridge : Royal Society of Chemistry, 2010. p. 209-242 (RSC Nanoscience & Nanotechnology; Vol. 15, No. 15).

Research output: Chapter in Book/Report/Conference proceedingChapterAcademicpeer-review

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T1 - Analysis of Proteins at the Single Cell Level

AU - le Gac, Severine

PY - 2010

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N2 - Proteins bring a consequent additional level of information in comparison with nucleic acids on a cell’s state as proteins are dynamically processed and chemically modified in the cell as a function of the cell life. Yet, the analysis of proteins is challenging as no amplification step is possible as is the case for nucleic acids, and another difficulty lies in the dynamic range of protein expression in a single sample (e.g. a single cell). While the same challenges are still found for microfluidic-based analysis of proteins, microsystems bring about enhanced analytical performance and novel analysis opportunities. This is illustrated here for two different strategies that can be adopted for protein analysis in a chip format. A first strategy consists of transposing the standard proteomic protocol in miniaturized analytical tools, and this provides a number of advantages and enhancement for the analysis: an overall improvement is expected when using smaller systems whose capacity matches better the size of the samples; sample manipulation is minimized when using LOC technology,and this goes together with a decrease in sample loss and contamination; enhanced analytical performance in terms of analysis time and detection sensitivity is ensured by micro- and nano-scale features; last, the use of microfabricated structures guarantees higher analysis reproducibility. In a second strategy, the analysis is actually performed at the single cell level. This strategy does not enable protein mapping anymore, but the investigation focuses on given proteins (a single protein of a small number thereof) which are specifically targeted. For that purpose, innovative microfluidic-based protocols have been developed, and we classify them in three categories of fully destructive, partially invasive and non invasive protocols. Ongoing developments in the area of nanotechnology would enable truly protein mapping at the single cell level, with the use of nanofabricated tools in a LOC platform.

AB - Proteins bring a consequent additional level of information in comparison with nucleic acids on a cell’s state as proteins are dynamically processed and chemically modified in the cell as a function of the cell life. Yet, the analysis of proteins is challenging as no amplification step is possible as is the case for nucleic acids, and another difficulty lies in the dynamic range of protein expression in a single sample (e.g. a single cell). While the same challenges are still found for microfluidic-based analysis of proteins, microsystems bring about enhanced analytical performance and novel analysis opportunities. This is illustrated here for two different strategies that can be adopted for protein analysis in a chip format. A first strategy consists of transposing the standard proteomic protocol in miniaturized analytical tools, and this provides a number of advantages and enhancement for the analysis: an overall improvement is expected when using smaller systems whose capacity matches better the size of the samples; sample manipulation is minimized when using LOC technology,and this goes together with a decrease in sample loss and contamination; enhanced analytical performance in terms of analysis time and detection sensitivity is ensured by micro- and nano-scale features; last, the use of microfabricated structures guarantees higher analysis reproducibility. In a second strategy, the analysis is actually performed at the single cell level. This strategy does not enable protein mapping anymore, but the investigation focuses on given proteins (a single protein of a small number thereof) which are specifically targeted. For that purpose, innovative microfluidic-based protocols have been developed, and we classify them in three categories of fully destructive, partially invasive and non invasive protocols. Ongoing developments in the area of nanotechnology would enable truly protein mapping at the single cell level, with the use of nanofabricated tools in a LOC platform.

KW - METIS-276315

KW - EWI-19430

KW - IR-75792

U2 - 10.1039/9781849732284-00209

DO - 10.1039/9781849732284-00209

M3 - Chapter

SN - 978-1-84755-911-1

T3 - RSC Nanoscience & Nanotechnology

SP - 209

EP - 242

BT - Unravelling Single Cell Genomics: Micro and Nanotools

A2 - Bontoux, Nathalie

A2 - Dauphinot, Luce

A2 - Potier, Marie-Claude

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le Gac S. Analysis of Proteins at the Single Cell Level. In Bontoux N, Dauphinot L, Potier M-C, editors, Unravelling Single Cell Genomics: Micro and Nanotools. Cambridge: Royal Society of Chemistry. 2010. p. 209-242. (RSC Nanoscience & Nanotechnology; 15). https://doi.org/10.1039/9781849732284-00209