Dual-Gradient Unified Chromatography: A New Paradigm for Versatility in Simultaneous Multicomponent Analysis.

Area of Science:

• Analytical Chemistry
• Chromatography
• Biopharmaceutical Analysis

Background:

• Analytical chemistry platforms are crucial for streamlining organic synthesis and biopharmaceutical processes.
• A need exists for versatile techniques capable of analyzing diverse molecules simultaneously.

Purpose of the Study:

• To introduce a novel hybrid separation technique, Dual-Gradient Unified Chromatography (DGUC).
• To demonstrate the capability of DGUC for simultaneous multicomponent analysis across a wide polarity range.
• To showcase DGUC's application in guiding the development of new (bio)pharmaceutical targets.

Main Methods:

• Development of DGUC based on automated dynamic modulation of CO2, organic modifier, and water blends with buffers.
• Investigation of fundamental aspects of the DGUC technique.
• Implementation of a DGUC-Diode Array Detector-Mass Spectrometry (DAD-MS) screening workflow with orthogonal column and mobile phase choices.

Main Results:

• DGUC enables simultaneous analysis of small and large molecules in a single experimental run.
• The technique covers a wide polarity range, offering a universal elution profile.
• Demonstrated analytical applications in guiding the development of pharmaceutical targets, including synthetic intermediates, nucleosides, peptides, proteins, and antibody-drug conjugates.

Conclusions:

• DGUC is a powerful and versatile hybrid separation technique for analytical chemistry.
• This framework significantly streamlines the analysis of complex (bio)pharmaceutical samples.
• DGUC offers a valuable tool for both academic and industrial research in drug discovery and development.