High Sensitivity Physicochemical Characterization of a Therapeutic Protein using the Agilent 1260 Infinity Bio-inert Quaternary LC with Agilent SEC and IEX Columns

In-depth physicochemical characterization of therapeutic proteins is required during all phases of drug development to ensure drug safety and efficacy. In this application note the new Agilent Infinity Bio-inert LC was used for peptide mapping, SEC and IEX of a therapeutic protein.

In this article we show results for size exclusion chromatography of Pl 28, a therapeutic protein under development at Gangagen, India. SEC can demonstrate protein integrity or prove the absence of dimers or multimer formation in its native conformation. A disadvantage of size exclusion is its lack of resolution. Small particle SEC columns produce superior resolution with minimal secondary interaction, providing a powerful tool for detecting impurities or multimer formation. Results were compared for two 5-�m particle columns and a 3-�m particle column. The second compound present in the mixture could only be detected with the Agilent Bio-SEC 3-�m.

The Agilent 1260 Infinity Bio-inert Quaternary LC system in combination with the Agilent Bio HPLC column portfolio provides a powerful and versatile tool for characterizing the physicochemical properties of the therapeutic protein, drug Pl 28.

The Agilent solution for analysis of therapeutic proteins achieves bio-inertness, superior resolution, corrosion resistance, high sensitivity and fast separation speed.

[Author Affiliation]

Katja Kornetzk, Agilent Technologies