Native HIC-MS of ADCs for DAR Species Determination and Identification

Discover how hydrophobic interaction chromatography interfaces in-line with mass spectrometry for ADC structural characterization and quantitation of drug-to-antibody ratios. This presentation highlights a recent publication on a novel HIC-MS approach that leverages the MS compatibility and kosmotropic ability of the thermally decomposable salt ammonium tartrate.

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Event Overview:

Antibody-drug conjugates (ADCs) have emerged as an attractive modality for cell- and tissue-specific delivery of therapeutic synthetic molecule payloads. One of the critical quality attributes of ADCs is the drug-to-antibody ratio (DAR), which significantly impacts their efficacy. Hydrophobic interaction chromatography (HIC) is routinely employed to determine the DAR of ADCs. However, the identification of ADC species from HIC separations poses challenges due to the incompatibility of traditional HIC methods, which use non–MS-compatible salts such as ammonium sulfate, with mass spectrometry (MS). Traditional identification workflows involve labor-intensive fractionation of HIC chromatographic peaks, followed by desalting and MS introduction through denaturing reversed-phase liquid chromatography (RPLC). This presentation highlights a recent publication on a novel HIC-MS approach that leverages the MS compatibility and kosmotropic ability of the thermally decomposable salt ammonium tartrate. This approach is enabled by directly coupling HIC with rapid size exclusion chromatography (SEC) for online desalting prior to native MS analysis with a simple instrument setup. The rapid SEC step efficiently separates much of the ammonium tartrate from the ADC species, enabling the characterization of both major and minor peaks from the HIC separation using MS. This workflow provides a straightforward method for native MS analysis of various DAR species in ADCs while maintaining excellent DAR species separation efficiency on HIC.

Key Learning Objectives:

• Introduce novel MS-compatible methods for native ADC structural characterization
• Highlight generalizable LC workflows for native-MS analysis of ADCs
• Review fundamentals of HIC and basic principles of ADC characterization

Who Should Attend:

• Antibody characterization scientists
• LC-MS development scientists
• Biopharma R&D scientists
• Analytical development chemists
• Quality control analysts
• Biotherapeutics discovery scientists

Speakers

Lance Cadang
Scientist
Genentech

Lance Cadang is an analytical chemist with over 8 years of experience at Genentech. He is a Scientist 3 in the Synthetic Molecule Analytical Chemistry group, where he serves as analytical lead for synthetic molecule programs and is responsible for developing analytical methods and control strategy. He develops and applies novel analytical methods and technologies to support the discovery, design, and CMC development of new therapeutic modalities, such as constrained peptides, macrocyclic peptides, oligonucleotides, and antibody-drug conjugates.

Corey Meadows, PhD
Commercial Product Manager
Tosoh Bioscience

Corey Meadows, PhD, obtained his PhD in physical chemistry at the University of California, Berkeley in 2014. He served as an analytical development scientist in the renewables and biopharmaceutical industries, with a focus on HPLC, LC-MS, and GC-MS techniques for small- and large-molecule analyses. While working within the biopharmaceutical industry, he focused primarily on antibody and protein therapeutic characterization using high-resolution LC-MS methods. He represents Tosoh Bioscience as a commercial product manager, focusing on application support and product development for analytical columns and multiangle light scattering (MALS) detectors..

Register Free: https://www.chromatographyonline.com/lcgc_w/hic-ms