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2. Bioanalysis of the Bicycle ® toxin conjugate BT5528 and released monomethyl auristatin E via liquid chromatography-tandem mass spectrometry.

Author

Tweed JA, Adams-Dam F, Allanson J, Holmes K, Senior R, Xu H, Li M, Bennett G, and Jeffrey P

Subjects
Humans, Chromatography, Liquid methods, Tandem Mass Spectrometry methods, Bicycling, Immunotoxins analysis, Immunoconjugates analysis, Antineoplastic Agents, Oligopeptides
Abstract

Background: The Bicycle ® toxin conjugate BT5528 is a novel peptide therapeutic conjugated to the cytotoxic agent monomethyl auristatin E (MMAE). A bioanalytical assay was developed to quantify BT5528 and unconjugated MMAE in human plasma. Methodology: BT5528 quantitation used a protein precipitation procedure followed by LC-MS/MS detection. Quantitation of MMAE required a selective offline and online solid-phase extraction with detection via LC-MS/MS. Results: BT5528 was quantified over the assay range of 5-2500 ng/ml and free MMAE was quantified over the assay range of 0.05-50 ng/ml. Conclusion: Bioanalytical methods were used in the bioanalysis of intact BT5528 and released MMAE, in a phase I/IIa clinical trial; to date, over 2000 human patient samples have been analyzed.

Published
2024
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