Recent mass spectrometry-based techniques and considerations for disulfide bond characterization in proteins

Area of Science:

• Protein chemistry
• Analytical chemistry
• Biopharmaceutical analysis

Background:

• Disulfide bonds are critical for protein structure, stability, and function.
• Biotherapeutics, especially monoclonal antibodies, rely heavily on disulfide bonds.
• Accurate disulfide bond analysis is essential for ensuring the safety and efficacy of protein therapeutics.

Purpose of the Study:

• To review recent mass spectrometry (MS)-based techniques for disulfide bond analysis in proteins.
• To provide guidance on efficient disulfide bond mapping strategies for biotherapeutics.
• To highlight considerations for sample preparation, fragmentation, and data analysis.

Main Methods:

• Focus on mass spectrometry (MS)-based techniques for disulfide bond characterization.
• Description of methods for sample preparation.
• Discussion of fragmentation techniques and automated algorithms for MS/MS data analysis.

Main Results:

• Recent MS-based techniques offer solutions for complex disulfide bond mapping.
• Automated algorithms facilitate rapid analysis of disulfide bonds from LC-MS/MS data.
• The review consolidates information on sample preparation, MS data, and software tools.

Conclusions:

• Mass spectrometry is the primary tool for disulfide bond analysis in proteins.
• Efficient disulfide bond mapping is vital for biotherapeutic development and quality control.
• This review serves as a resource for researchers developing new methods and characterizing biologics.