# Accelerating Characterization of Therapeutic Antibodies: A Comparative Assessment of icIEF-UV/MS and the Traditional Fractionation Workflow

## TL;DR

This paper compares a new method for analyzing antibody variants using icIEF-UV/MS with traditional methods, showing it is faster and more efficient.

## Contribution

The study introduces a faster and more efficient method for characterizing therapeutic antibody variants using icIEF-UV/MS.

## Key findings

- icIEF-UV/MS provides comparable separation and quantitation to traditional methods.
- icIEF-UV/MS offers superior direct MS-based peak characterization with less time and sample processing.
- The new method is more efficient for analyzing charge variants in therapeutic antibodies.

## Abstract

Accurate characterization of charge variants in biologics
is crucial
to uphold stringent quality standards ensuring the safety and efficacy
of biotherapeutic products and regulatory requirements for the Biologic
Licensing Application (BLA) process enabling marketing application.
These variants, arising from post-translational modifications (PTMs)
during upstream processing and enzymatic/nonenzymatic reactions in
downstream processing and storage, can significantly impact therapeutic
potency, efficacy, and immunogenicity. Conventional methods for characterizing
charge variants typically involve labor-intensive fraction enrichment,
consuming time and resources. However, recent technological advancements,
exemplified by the Intabio ZT system’s innovative platform,
enable the seamless and direct integration of imaged capillary isoelectric
focusing (icIEF) and UV quantitation with mass spectrometry (MS) PTM
identification, facilitating rapid and unbiased characterization.
In this study, we conduct a comparative assessment of the charge variant
characterization of investigative mAb-1 using icIEF-UV/MS analysis
and a traditional fractionation-based workflow. Our results demonstrated
that icIEF-UV/MS-based proteoform characterization provided comparable
icIEF-UV separation and quantitation and superior direct MS-based
peak characterization with shorter time and less sample processing
compared to conventional fractionation approaches.