Fully automated 2D-LC/MS for biotherapeutics charge variant characterization using Ion Exchange (IEX) and Reversed-Phase (RP) Chromatography

During the manufacturing process, protein therapeutics (mAbs) are subjected to several posttranslational modifications (PTMs) which will affect the biochemical and biophysical properties of the mAbs. One of such events is charge variants and its determination is one of the most important quality attributes for the characterization of biopharmaceuticals such as monoclonal antibodies (mAbs). Traditionally, charge variant profiling is done using ion exchange chromatography (IEX), fractionation collection, followed offline desalting and LC-MS analysis to qualify the charge variant pattern of the mAbs. This approach is not a straightforward procedure, mostly due to the incompatibility of the IEX buffers (phosphate buffers containing sodium chloride) with the electrospray ionization process.

However, a 2D-LC solution enables automated desalting, and offers additional separation by the reversed-phase chromatography (RP) in the second dimension.

In this webinar, the speakers will discuss the importance of charge variants analysis of therapeutic mAbs and a fully automated multiple heart-cutting 2D-LC/MS workflow solution for charge variant characterization.

Presenter: Prof. Anurag S. Rathore (Department of Chemical Engineering, Indian Institute of Technology, Delhi)

Presenter: Ms. Paramjeet Khandpur (Product Specialist, Agilent Techologies, India)

Presenter: Dr. Ashish Pargaonkar (Senior Applications Scientist, Agilent Technologies, India)