High Resolution LC/MS Separation and Characterization of Chemoenzymatic Site-specific Engineered Antibody-Drug Conjugates

[Orbitrap_SciLib]

Shanhua Lin1, Terry Zhang2, Jonathan Josephs2, Brian Agnew3, and Xiaodong Liu1
ASMS 2017 Poster
Purpose: Demonstrate application of the SiteClick™ technology to the production of antibody-drug conjugates (ADCs) and a number of different detection probes. Methods: ADCs were made from SiteClick™ enzyme-based N-glycan labeling of bevacizumab and panitumumab with DIBO-MMAE and DIBO-DFO. F(ab’)2 and scFc fragments from unconjugated, azide-tagged intermediate, and ADCs were analyzed using a Thermo Scientific™ MAbPac™ RP column coupled with a Thermo Scientific™ Orbitrap™ Fusion™ mass spectrometer. Results: Enzymatic pre-treatment, or “trimming” of the antibody glycans with GlycINATOR® endoglycosidase S2, prior to azide activation, results in highly-reproducible degrees of labeling (DOLs) of 2.0.


1. Thermo Fisher Scientific, Sunnyvale, CA 2. Thermo Fisher Scientific, San Jose, CA 3. Thermo Fisher Scientific, Eugene, OR