Automated multi-attribute method sample preparation using high-throughput buffer exchange tips

Area of Science:

• Biopharmaceutical analysis
• Mass spectrometry
• Analytical chemistry

Background:

• The multi-attribute method (MAM) is a key mass spectrometry (MS)-based technique for characterizing biotherapeutics like monoclonal antibodies (mAbs).
• Enhancing sample preparation throughput is crucial for expanding MAM applications in drug development without compromising data quality.

Purpose of the Study:

• To develop and validate a fully automated sample preparation protocol for MAM analysis.
• To improve the speed and efficiency of sample preparation for biotherapeutic characterization.

Main Methods:

• A fully automated protocol utilizing miniaturized size-exclusion chromatography columns in pipette tips for rapid desalting (<15 minutes).
• Sample analysis via electrospray ionization (ESI) orbitrap mass spectrometry coupled with ultra-high-performance liquid chromatography (UHPLC) and dual column switching.

Main Results:

• Automated sample preparation showed no significant changes in product attributes or quantities compared to manual methods.
• Enhanced sample recovery and excellent reproducibility were achieved across various sample concentrations.
• Individual columns offered flexibility in sample batch size and plate location, unlike plate-based systems.

Conclusions:

• The automated MAM protocol significantly reduces "at-bench" time for preparing up to 96 samples.
• This automation facilitates process development and routine quality control applications of MAM.
• The method provides a robust and efficient solution for high-throughput biotherapeutic characterization.