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Updated: Oct 5, 2025
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Characterizing Individual Protein Aggregates by Infrared Nanospectroscopy and Atomic Force Microscopy
Published on: September 12, 2019
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Methods for the Characterization of Protein Aggregates.
Marc Martínez-Miguel1, Witold Tatkiewicz1, Mariana Köber1
1Institut de Ciència de Materials de Barcelona, ICMAB-CSIC and CIBER de Bioingeniería, Biomateriales y Nanomedicina (CIBER-BBN), Campus UAB, Bellaterra, Spain.
Methods in Molecular Biology (Clifton, N.J.)
|January 28, 2022
Summary
Characterizing protein aggregates, like bacterial inclusion bodies (IBs), is crucial for understanding diseases. This study details methods for analyzing their physicochemical and nanoscale properties, aiding in their application as biomaterials.
Area of Science:
- Biophysics
- Materials Science
- Nanotechnology
Background:
- Protein aggregates are hallmarks of various diseases.
- Bacterial inclusion bodies (IBs) are pure, nanoscale protein aggregates with potential applications.
- Limited knowledge exists regarding the physicochemical properties of IBs.
Purpose of the Study:
- To present methods for characterizing protein aggregates as particulate materials.
- To detail techniques for assessing physicochemical and nanoscale properties of protein aggregates.
- To highlight the utility of these methods for understanding protein aggregate behavior and applications.
Main Methods:
- Dynamic Light Scattering (DLS) for sizing.
- Nanoparticle Tracking Analysis (NTA) for sizing and counting.
- Zeta potential for colloidal stability.
- Atomic Force Microscopy (AFM) and Scanning Electron Microscopy (SEM) for morphology.
- Cryo-Transmission Electron Microscopy (cryo-TEM) for internal structure.
- Contact Angle (CA) for wettability.
- Force Spectroscopic AFM (FS-AFM) for nanomechanical properties.
- 4'4-dithiodipyridine (DTDP) method for accessible sulfhydryl groups.
Main Results:
- The chapter provides a comprehensive overview of established and advanced techniques for protein aggregate characterization.
- Detailed descriptions of methods enable the assessment of size, stability, morphology, internal structure, wettability, mechanical properties, and chemical composition.
- Examples illustrate the capabilities and applications of each technique in analyzing protein aggregates.
Conclusions:
- A robust set of methods exists for the comprehensive physicochemical and nanoscale characterization of protein aggregates.
- Understanding these properties is essential for both fundamental research into diseases and the development of engineered protein-based biomaterials.
- This chapter serves as a guide for researchers utilizing these techniques for protein aggregate analysis.
