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Multiple cellular antigen detection by ICP-MS

O Ornatsky1, V I Baranov, D R Bandura

  • 1Institute for Biomaterials and Biomedical Engineering, University of Toronto, 4 Taddlecreek Rd., Rm. 407, Toronto, ON, Canada, M5S 3G9. olga.ornatsky@utoronto.ca

Journal of Immunological Methods
|December 13, 2005
PubMed

Insights

A new ICP-MS-linked metal-tagged immunophenotyping method allows simultaneous detection of multiple proteins in single cells, overcoming limitations of current fluorescence methods for multiplexed proteomic analysis.

Area of Science:

  • Cell biology
  • Proteomics
  • Immunophenotyping

Background:

  • Simultaneous detection of multiple intracellular and extracellular proteins in single cells is crucial in cell biology.
  • Current fluorescence-activated flow cytometry methods have limitations in multiplexing capabilities.
  • There is a need for advanced techniques for high-level multiplexed proteomic analysis.

Purpose of the Study:

  • To develop and evaluate a novel ICP-MS-linked metal-tagged immunophenotyping method for highly multiplexed proteomic analysis.
  • To assess the feasibility of simultaneous detection of multiple protein targets within single cells.
  • To compare the performance of ICP-MS-based immunoassays with traditional FACS methods.

Main Methods:

  • Developed a novel ICP-MS-linked metal-tagged immunophenotyping technique.
  • Utilized metal tags (Au, Sm, Eu, Tb) conjugated to secondary antibodies for multiplexed detection.
  • Investigated the expression of BCR/Abl, CD33, c-Kit, and VLA-4 in human leukemia cell lines.
  • Compared ICP-MS results with data obtained from Fluorescence-Activated Cell Sorting (FACS).

Main Results:

  • Successfully demonstrated the simultaneous detection of four antigens (4-plex assay) using metal-tagged antibodies and ICP-MS.
  • ICP-MS offers several advantages over traditional methods for enhancing immunoassay performance.
  • Preliminary findings suggest the potential for detecting more than four antigens simultaneously with ICP-MS.

Conclusions:

  • ICP-MS-linked metal-tagged immunophenotyping is a promising approach for highly multiplexed proteomic analysis in cell biology.
  • This novel method overcomes the multiplexing limitations of current optical techniques.
  • Further research and feasibility studies are expected to expand the multiplexing capacity of this technique.

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