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Determination of Antibody–Drug Conjugate Released Payload Species Using Directed in Vitro Assays and Mass Spectrometric Interrogation
journal contribution
posted on 2016-05-20, 00:00 authored by Andrew J. Bessire, T. Eric Ballard, Manoj Charati, Justin Cohen, Michael Green, My-Hanh Lam, Frank Loganzo, Birte Nolting, Betsy Pierce, Sujiet Puthenveetil, Lee Roberts, Klaas Schildknegt, Chakrapani SubramanyamAntibody-drug conjugates
(ADC) are currently an active area of
research, focused primarily on oncology therapeutics, but also to
a limited extent on other areas such as infectious disease. The success
of this type of targeted drug delivery is dependent upon many factors,
one of which is the performance of the linker in releasing an active
drug moiety under the appropriate conditions. As a tool in the development
of linker/payload chemistry, we have developed an in vitro method
for the identification of payload species released from ADCs in the
presence of lysosomal enzymes. This method utilizes commercially available
human liver S9 fraction as the source of these enzymes, and this has
certain advantages over lysosomal fractions or purified enzymes. This
article describes the characterization and performance of this assay
with multiple ADCs composed of known and novel linkers and payloads.
Additionally, we report the observation of incomplete degradation
of mAb protein chains by lysosomal enzymes in vitro, believed to be
the first report of this phenomenon involving an ADC therapeutic.
