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Proton Transfer and Protein Conformation Dynamics in Photosensitive Proteins by Time-resolved Step-scan Fourier-transform Infrared Spectroscopy
Published on: June 27, 2014
Infrared spectroscopy for biopharmaceutical protein analysis
Peter Christian Gross1, Michael Zeppezauer
1Symbiotec GmbH, Science Park Saar 1, D-66123 Saarbrücken, Germany.
Journal of Pharmaceutical and Biomedical Analysis
|April 13, 2010
Summary
A new mid-infrared transmission spectroscopy method offers high-throughput protein analysis for biopharmaceutical quality control. This technique provides quantitative analysis, identity and purity determination, and structural characterization of protein therapeutics.
Area of Science:
- Biotechnology
- Analytical Chemistry
- Spectroscopy
Background:
- Current methods for protein therapeutic analysis, such as attenuated total reflection (ATR), are often surface-sensitive.
- There is a need for robust, quantitative, and high-throughput analytical techniques for biopharmaceutical quality control.
Purpose of the Study:
- To develop and validate a mid-infrared transmission spectroscopic method for protein therapeutics.
- To compare the advantages of transmission spectroscopy with existing methods like ATR.
- To demonstrate the method's utility in characterizing protein structure, purity, and interactions.
Main Methods:
- Development and validation of a mid-infrared transmission spectroscopic method.
- Application of Lambert-Beer's law for quantitative analysis.
- Establishment of specific identity criteria for purity determination.
- Characterization of protein structure, aggregation, and ligand binding in solution.
Main Results:
- The transmission method demonstrated high sample throughput and compliance with Good Manufacturing Practice (GMP) requirements.
- Advantages over ATR include quantitative analysis, identity and purity determination, structural characterization, and ligand binding studies.
- Successful characterization of ONCOHIST drug substance and its interaction with phosphate ions was achieved.
Conclusions:
- Mid-infrared transmission spectroscopy is a powerful and versatile tool for protein analysis in biotechnology.
- This method supplements existing analytical techniques for biopharmaceutical quality control.
- The developed method offers significant advantages for the comprehensive analysis of therapeutic proteins.
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