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

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.

Bioanalysis of Targeted Nanoparticles in Monkey Plasma via LC-MS/MS.

This work represents the first reporting of a comprehensive bioanalytical GLP methodology detailing the mass spectrometric quantitation of PF-05212384 dosed as a targeted polymeric encapsulated nanoparticle (PF-07034663) to monkeys. Polymeric nanoparticles are a type of drug formulation that enables the sustained release of an active therapeutic agent (payload) for targeted delivery to specific sites of action such as cancer cells. Through the careful design and engineering of the nanoparticle formulation, it is possible to improve the biodistribution and safety of a given therapeutic payload in circulation. However, the bioanalysis of nanoparticles is challenging due to the complexity of the nanoparticle drug formulation itself and the number of pharmacokinetic end points needed to characterize the in vivo exposure of the nanoparticles. Gedatolisib, also known as PF-05212384, was reformulated as an encapsulated targeted polymeric nanoparticle. The bioanalytical assays were validated to quantitate both total and released PF-05212384 derived from the encapsulated nanoparticle (PF-07034663). Assay performance calculated from quality control samples in three batch runs demonstrated intraday precision and accuracy within 10.3 and 12.2%, respectively, and interday precision and accuracy within 9.1 and 8.5%, respectively. This method leveraged automation to ease the burden of a laborious and complicated sample pretreatment and extraction procedure. The automated method was used to support a preclinical safety study in monkeys in which both released and total PF-05212384 concentrations were determined in over 1600 monkey plasma study samples via LC-MS/MS.

Boston Society's 11th Annual Applied Pharmaceutical Analysis conference.

Boston Society's 11th Annual Applied Pharmaceutical Analysis conference, Hyatt Regency Hotel, Cambridge, MA, USA, 14-16 September 2015 The Boston Society's 11th Annual Applied Pharmaceutical Analysis (APA) conference took place at the Hyatt Regency hotel in Cambridge, MA, on 14-16 September 2015. The 3-day conference affords pharmaceutical professionals, academic researchers and industry regulators the opportunity to collectively participate in meaningful and relevant discussions impacting the areas of pharmaceutical drug development. The APA conference was organized in three workshops encompassing the disciplines of regulated bioanalysis, discovery bioanalysis (encompassing new and emerging technologies) and biotransformation. The conference included a short course titled 'Bioanalytical considerations for the clinical development of antibody-drug conjugates (ADCs)', an engaging poster session, several panel and round table discussions and over 50 diverse talks from leading industry and academic scientists.

Automated supported liquid extraction using 2D barcode processing for routine toxicokinetic portfolio support.

BACKGROUND: A new bioanalytical sample preparation approach has been developed to enhance the efficiency, reduce errors and improve the data quality supporting routine toxicokinetic (TK) study samples analysis, via the implementation of 2D barcode processing coupled with fully automated supported liquid extraction (SLE). RESULTS: A fully automated SLE was validated and used to determine TK drug concentrations of over 500 unknown samples via 2D barcode processing. Assay performance calculated from a total of 291 quality control samples over the period of validation through sample analysis demonstrated inter-day precision and accuracy within 10 and 7.3%, respectively. CONCLUSION: A new logistical approach implementing the use of 2D barcodes and automated SLE demonstrates the potential of a new methodology for the routine bioanalytical support of TK study sample analysis.

Automated sample preparation for regulated bioanalysis: an integrated multiple assay extraction platform using robotic liquid handling.

BACKGROUND: A novel approach for regulated bioanalytical sample preparation has been developed to combine multiple types of extraction techniques into one integrated and automated sample-preparation suite that pairs a graphical user interface with the Hamilton Microlab(®) STAR robotic liquid handler. RESULTS: The multi-assay sample-preparation suite is composed of three bioanalytical extraction techniques: protein precipitation, solid-phase extraction and liquid-liquid extraction. Validation data provided highly reproducible and robust results for each respective automated extraction technique. CONCLUSION: The user-friendly graphical user interface and modular method design provide a flexible and versatile approach for routine bioanalytical sample-preparation and is the first fully integrated multiple assay sample-preparation suite for regulated bioanalysis.