FabRICATOR®-HPLC for Automated Antibody Subunit Analysis

December 17, 2019 | References |

Middle-level analysis is a universally accepted analytical strategy for rapid characterization of antibodies. The FabRICATOR® (IdeS)enzyme specifically digests IgG below the hinge, generating  fragments that are suitable for high-resolution mass spectrometry. By immobilizing the FabRICATOR enzyme in an HPLC column format, a fully automated on-column digestion of IgG is possible.

In a recent article written by the Genovis team and published in Chromatography Today, an LC-MS workflow for automated middle-level analysis of monoclonal antibodies (mAbs) and mAbs-based biologics is described and evaluated. FabRICATOR-HPLC was shown to facilitate easy on-line sample preparation for middle-level LC-MS. On-column digestion of mAbs followed by on-column reduction and analysis by RP-HPLC and MS yielded results indistinguishable from those produced using an off-line sample preparation protocol.

“With minimal carry-over, low sample requirements and a tolerance for a wide concentration range, this method is very versatile. It is well suited for both routine analysis as well as more advanced applications such as automatic monitoring of mAb CQAs during production in a bioreactor.”

The full article is available here:

Nägeli et al., 2020. On-column digestion of mAbs for automated middle-level analysis by LC-MS. Chromatograpy Today

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