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Related Concept Videos

Proteomics01:33

Proteomics

8.6K
A proteome is the entire set of proteins that a cell type produces. We can study proteomes using the knowledge of genomes because genes code for mRNAs, and the mRNAs encode proteins. Although mRNA analysis is a step in the right direction, not all mRNAs are translated into proteins.
Proteomics is the study of proteomes' function. It involves the large-scale systematic study of the proteome to denote the protein complement expressed by a genome. Scientist Mark Wilkins coined the term...
8.6K

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Nanosensors to Detect Protease Activity In Vivo for Noninvasive Diagnostics
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Nanoparticle-Aided Nanoreactor for Nanoproteomics.

Zhichang Yang1, Zhaoran Zhang2, Daoyang Chen1

  • 1Department of Chemistry, Michigan State University, 578 S Shaw Lane, East Lansing, Michigan 48824 United States.

Analytical Chemistry
|July 23, 2021
PubMed
Summary

A new nanoparticle-aided nanoreactor (Nano3) technique significantly enhances protein identification from limited cell samples. This method improves proteome profiling for few-cell and single-cell studies, advancing research in cell heterogeneity.

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Fast Enzymatic Processing of Proteins for MS Detection with a Flow-through Microreactor
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Rapid, Scalable Assembly and Loading of Bioactive Proteins and Immunostimulants into Diverse Synthetic Nanocarriers Via Flash Nanoprecipitation
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Area of Science:

  • Proteomics
  • Cell Biology
  • Biochemistry

Background:

  • Understanding cell-to-cell heterogeneity is vital for disease and developmental research.
  • Few-cell and single-cell proteomics require highly sensitive methodologies.
  • Efficient sample processing with high recovery and minimal contamination is critical.

Purpose of the Study:

  • To develop a novel, highly sensitive proteomic technique for analyzing minute amounts of mammalian cell proteins.
  • To improve sample processing for low-nanogram quantities, enabling better proteome profiling.
  • To establish a method for effective bottom-up proteomics on a small number of cells.

Main Methods:

  • Developed the nanoparticle-aided nanoreactor for nanoproteomics (Nano3) technique.
  • Utilized a nanoreactor (≤30 nL) with packed nanoparticles for protein concentration, cleaning, and digestion.
  • Integrated Nano3 with nanoRPLC-MS/MS for proteome profiling of low-nanogram protein samples and 10-1000 HeLa cells.

Main Results:

  • The Nano3 method identified 40 times more proteins from 2-ng mouse brain samples compared to the SP3 method.
  • Nano3 demonstrated significantly higher sample recovery for mass-limited proteome samples.
  • Processing 10 HeLa cells yielded 441 (MS/MS) and 983 (MBR+MS/MS) protein identifications.

Conclusions:

  • The Nano3 technique offers a substantial improvement in protein identification sensitivity for low-input proteomics.
  • This method is effective for processing few mammalian cells, facilitating detailed proteome profiling.
  • Nano3 represents a valuable advancement for few-cell and single-cell proteomics research.