Accelerating biotherapeutics discovery: Blueprint for a universal LC-MALDI-MS offline approach

Area of Science:

• Biopharmaceutical analysis
• Analytical chemistry
• Mass spectrometry

Background:

• Biotherapeutics are crucial for treating various diseases.
• High-throughput liquid chromatography (LC) is vital for biotherapeutic discovery.
• Traditional LC methods face challenges with mass spectrometry (MS) compatibility, slowing down impurity identification.

Purpose of the Study:

• To develop a rapid and automated workflow for biotherapeutic peak identification.
• To overcome MS compatibility issues with common LC techniques.
• To accelerate the discovery and development of next-generation biologics.

Main Methods:

• Developed an offline LC-MALDI-MS workflow using an LC-fraction collector and MALDI-TOF-MS.
• The approach is compatible with various LC techniques, including size exclusion chromatography (SEC), hydrophobic interaction chromatography (HIC), and Reversed Phase LC.
• The workflow is designed for automation and minimal sample preparation.

Main Results:

• Achieved rapid peak identification within hours.
• Demonstrated versatility across different chromatographic methods and analytes (antibodies, fusion proteins, peptides).
• Overcame buffer and matrix interferences often seen with electrospray ionization (ESI-MS).

Conclusions:

• The automated LC-MALDI-MS workflow significantly streamlines biotherapeutic impurity identification.
• This method accelerates drug discovery timelines by enabling rapid analysis.
• The workflow is broadly applicable to various biopharmaceutical development stages.