High-Speed Automated Microdroplet Reactions with Ion-Mobility for Rapid Therapeutic Protein Characterization.

Area of Science:

• Biochemistry and Biophysics
• Analytical Chemistry
• Protein Science

Background:

• Traditional protein characterization methods require large sample amounts (10-20 μg) and can introduce artifacts.
• Post-translational modifications (PTMs) complicate the accurate characterization of protein therapeutics.
• Rapid and precise characterization is crucial for efficient protein therapeutic discovery.

Purpose of the Study:

• To develop an ultrafast, microdroplet-based reaction method for protein digestion and reduction.
• To establish a rapid and reliable workflow for protein therapeutic characterization.
• To integrate ion mobility separation with microdroplet reactions for enhanced mass species analysis.

Main Methods:

• Automated microdroplet-based reactions for protein digestion and reduction in under one millisecond.
• Integration of ion mobility separation with microdroplet reactions.
• Application across diverse therapeutic protein formats including IgG1 antibodies, Fc-cytokine fusion proteins, and antibody-drug conjugates (ADCs).

Main Results:

• Demonstrated ultrafast protein digestion and reduction (<1 ms).
• Achieved enhanced throughput in protein therapeutic characterization without compromising accuracy.
• Successfully applied the technique to various therapeutic protein modalities.

Conclusions:

• The developed microdroplet reaction technique offers a high-throughput workflow for comprehensive protein analysis.
• This innovative approach accelerates the characterization of protein therapeutics, aiding early-stage discovery.
• First report of online microdroplet reactions integrated with ion mobility separation across multiple therapeutic modalities.