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

Yuko Ogata¹, Pamela M Quizon², Nancy S Nightlinger¹

¹Just-Evotec Biologics, Inc., Seattle, WA, USA.

Rapid Communications in Mass Spectrometry : RCM

|November 16, 2021

Summary

An automated multi-attribute method (MAM) sample preparation protocol significantly enhances throughput for biotherapeutic analysis. This method achieves rapid desalting and maintains product attribute integrity, supporting process development and quality control.

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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.