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Updated: Jun 24, 2026

How to Stabilize Protein: Stability Screens for Thermal Shift Assays and Nano Differential Scanning Fluorimetry in the Virus-X Project

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Published on: February 11, 2019

Infrared techniques for quantifying protein structural stability.

John S Vrettos¹, Curtis W Meuse

¹Biochemical Science Division, Chemical Science and Technology Laboratory, National Institute of Standards and Technology, 100 Bureau Drive Stop 8313, Gaithersburg, MD 20899-8313, USA.

Analytical Biochemistry

|March 31, 2009

Summary

Infrared spectroscopy quantifies protein structural stability using amide hydrogen/deuterium exchange. This method effectively assesses protein interactions and stability in various states for biopharmaceutical applications.

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Characterizing Individual Protein Aggregates by Infrared Nanospectroscopy and Atomic Force Microscopy

Characterizing Individual Protein Aggregates by Infrared Nanospectroscopy and Atomic Force Microscopy

Published on: September 12, 2019

Area of Science:

Biophysical Chemistry
Structural Biology
Spectroscopy

Background:

Biopharmaceutical and regulatory sectors need robust methods to assess protein structural stability and interactions.
Current methods may have limitations regarding sample amount, molecular weight, modifications, or sample environment.
Infrared spectroscopy offers a sensitive, versatile alternative for analyzing proteins in diverse states.

Purpose of the Study:

To investigate and validate infrared spectroscopy for quantifying protein structural stability.
To measure the extent of amide hydrogen/deuterium exchange for assessing stability.
To apply the method to proteins in solution, bound to ligands, and interacting with lipid membranes.

Main Methods:

Utilized infrared spectroscopy to monitor amide hydrogen/deuterium exchange in proteins.
Applied the technique to proteins in solution and complexed with ligands or lipid membranes.
Measured thermodynamic stability of lysozyme, trypsin-inhibitor complexes, and cytochrome c-lipid membrane interactions.

Main Results:

Demonstrated infrared spectroscopy's capability to quantify protein structural stability.
Successfully measured amide hydrogen/deuterium exchange for various protein systems.
Reported thermodynamic stability data for diverse protein preparations, including membrane interactions.

Conclusions:

Infrared spectroscopy is a powerful tool for assessing protein structural stability.
The method is applicable to a wide range of protein states and interactions.
This technique provides valuable data for biopharmaceutical and biotechnology research and development.