Label-Free Quantification of Molecular Interaction in Live Red Blood Cells by Tracking Nanometer Scale Membrane Fluctuations

Bo Yao, Yunze Yang, Nanxi Yu, Nongjian Tao, Di Wang, Shaopeng Wang, Fenni Zhang

Research output: Contribution to journalArticlepeer-review

Abstract

Molecular interactions in live cells play an important role in both cellular functions and drug discovery. Current methods for measuring binding kinetics involve extracting the membrane protein and labeling, while the in situ quantification of molecular interaction with surface plasmon resonance (SPR) imaging mainly worked with fixed cells due to the micro-motion related noises of live cells. Here, an optical imaging method is presented to measure the molecular interaction with live red blood cells by tracking the nanometer membrane fluctuations. The membrane fluctuation dynamics are measured by tracking the membrane displacement during glycoprotein interaction. The data are analyzed with a thermodynamic model to determine the elastic properties of the cell observing reduced membrane fluctuations under fixatives, indicating cell fixations affect membrane mechanical properties. The binding kinetics of glycoprotein to several lectins are obtained by tracking the membrane fluctuation amplitude changes on single live cells. The binding kinetics and strength of different lectins are quite different, indicating the glycoproteins expression heterogeneity in single cells. It is anticipated that the method will contribute to the understanding of mechanisms of cell interaction and communication, and have potential applications in the mechanical assessment of cancer or other diseases at the single-cell level, and screening of membrane protein targeting drugs.

Original languageEnglish (US)
Article number2201623
JournalSmall
Volume18
Issue number28
DOIs
StatePublished - Jul 14 2022
Externally publishedYes

Keywords

  • binding kinetics
  • lectin
  • live red blood cells
  • membrane fluctuation
  • optical imaging

ASJC Scopus subject areas

  • Biotechnology
  • Chemistry(all)
  • Biomaterials
  • Materials Science(all)

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