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Collision-Based Electrochemical Detection of Lysozyme Aggregation.

Kevin A Kirk1, Alina Vasilescu2, Daniel Andreescu1

  • 1Department of Chemistry and Biomolecular Science, Clarkson University, Potsdam, New York 13699-5810, United States.

Analytical Chemistry
|January 8, 2021
PubMed
Summary

Single-particle collision electrochemistry (SPCE) using silver nanoparticle (AgNP) probes offers a novel method for tracking protein aggregation. This technique monitors AgNP collisions to assess lysozyme (Lyz) aggregation states in solution.

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Area of Science:

  • Biophysical Chemistry
  • Nanotechnology
  • Electrochemistry

Background:

  • Protein aggregation is critical in pharmaceutical and biomedical industries, necessitating accurate monitoring methods.
  • Traditional techniques like AFM and electron microscopy have limitations, including sample pre-treatment and lack of native state representation.
  • Developing solution-based, rapid, and accurate methods for protein aggregation analysis is essential for quality control.

Purpose of the Study:

  • To introduce and validate single-particle collision electrochemistry (SPCE) as a novel technique for early detection and tracking of protein aggregation.
  • To demonstrate the utility of SPCE using silver nanoparticle (AgNP) redox probes for monitoring lysozyme (Lyz) aggregation.
  • To establish SPCE as a viable alternative to traditional methods for protein stability assessment.

Main Methods:

  • Utilizing single-particle collision electrochemistry (SPCE) with silver nanoparticle (AgNP) redox probes to monitor protein-analyte interactions.
  • Tracking the frequency of oxidative impacts of AgNPs in a chronoamperometric profile, which is influenced by protein aggregation.
  • Analyzing the interaction of lysozyme (Lyz) with AgNPs, observing changes in AgNP surface coverage and conjugate formation based on aggregation state.

Main Results:

  • SPCE effectively monitored lysozyme aggregation by detecting changes in the frequency of AgNP collisions.
  • Non-aggregated lysozyme stabilized AgNPs, while aggregated lysozyme promoted conjugate agglomeration, altering collision frequency.
  • Electrochemical data showed statistically significant differences (99% confidence) in charge transfer between aggregated and non-aggregated lysozyme states.
  • Results correlated well with UV-vis, circular dichroism, and AFM, validating SPCE's efficacy.

Conclusions:

  • Single-particle collision electrochemistry (SPCE) provides a sensitive and accurate method for tracking protein aggregation in real-time.
  • This electrochemical technique offers a powerful tool for indirectly evaluating protein stability and screening for aggregate formation.
  • SPCE presents a promising alternative for protein analysis, overcoming limitations of traditional characterization methods.