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Related Experiment Video

Updated: May 10, 2026

Flow Cytometric Analysis of Bimolecular Fluorescence Complementation: A High Throughput Quantitative Method to Study Protein-protein Interaction
11:11

Flow Cytometric Analysis of Bimolecular Fluorescence Complementation: A High Throughput Quantitative Method to Study Protein-protein Interaction

Published on: August 15, 2013

An on-bacterium flow cytometric immunoassay for protein quantification.

Wen-Jun Lan1, Wei Lan, Hai-Yan Wang

  • 1Institute of Biomedical Engineering & Shandong Provincial Key Laboratory of Microbial Engineering, Shandong Institute of Light Industry, Jinan 250353, China. lanwenjun0522@sina.com

Journal of Pharmaceutical and Biomedical Analysis
|June 7, 2013
PubMed
Summary

This study introduces an on-bacterium flow cytometric immunoassay using Staphylococcus aureus (SAC) as a bead alternative for protein quantification. This novel method offers a simpler, more convenient approach for detecting biomarkers like CEA and CYFRA 21-1.

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Area of Science:

  • Biotechnology
  • Immunology
  • Analytical Chemistry

Background:

  • Polystyrene bead-based flow cytometric immunoassays are common but require inconvenient bead preparation.
  • There is a need for simpler, more accessible methods for protein quantification.

Purpose of the Study:

  • To develop and validate a simple, on-bacterium flow cytometric immunoassay for protein quantification.
  • To utilize Staphylococcus aureus (SAC) as a carrier to replace traditional polystyrene beads.

Main Methods:

  • SAC beads were prepared via CFSE labeling, paraformaldehyde fixation, and antibody binding.
  • The assay was validated using Carcinoembryonic antigen (CEA) and cytokeratin-19 fragment (CYFRA 21-1) as model proteins.
  • Quantification was performed using biotinylated proteins and streptavidin-phycoerythrin (SA-PE) with flow cytometry.

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Last Updated: May 10, 2026

Flow Cytometric Analysis of Bimolecular Fluorescence Complementation: A High Throughput Quantitative Method to Study Protein-protein Interaction
11:11

Flow Cytometric Analysis of Bimolecular Fluorescence Complementation: A High Throughput Quantitative Method to Study Protein-protein Interaction

Published on: August 15, 2013

Quantitative Immunofluorescence Assay to Measure the Variation in Protein Levels at Centrosomes
09:39

Quantitative Immunofluorescence Assay to Measure the Variation in Protein Levels at Centrosomes

Published on: December 20, 2014

IP-FCM: Immunoprecipitation Detected by Flow Cytometry
12:17

IP-FCM: Immunoprecipitation Detected by Flow Cytometry

Published on: December 2, 2010

Main Results:

  • A strong correlation was observed between protein concentration and PE mean fluorescence intensity (MFI).
  • The immunoassay achieved a limit of detection (LOD) of at least 0.25 ng/ml.
  • High precision and accuracy were demonstrated, with relative standard deviation (R.S.D.) and relative error (R.E.) below 10%.

Conclusions:

  • The on-bacterium flow cytometric immunoassay provides a convenient and effective alternative to polystyrene bead-based methods.
  • This assay shows high correlation with existing methods for serum protein quantification (CEA and CYFRA 21-1).
  • The developed method holds potential for routine serum protein quantification.