The CSSC And CTMSDG Invite You To The November Discussion Group Meeting On November 19, 2015 Featuring Two Speakers!
Cong Wei, PhD, Pfizer: Profiling ADME attributes of Antibody-Drug Conjugates
Mary Joan Castillo, UCONN: Targeted quantitative proteomics for cancer biomarker validation via high
sample throughput N-in-1 uMS
WHERE: Sheraton Four Points Hotel, Meriden CT Link
WHEN: Registration: 5pm Dinner: 5:30-6:30pm Presentations: 6:30-8 Pm
COST: Free
Cong Wei, PhD
Cong Wei earned her Ph.D. degree of Pharmacology from University of Pennsylvania. Her graduate work focused on identification and characterization of lipid metabolites from cancer cells by LC-MS. After graduation in 2010, she worked for Bristol-Myers Squibb in the Bioanalytical and Discovery Analytical Department for almost four years. Her work at BMS included method development and sample analysis on both endogenous and exogenous small molecules and big molecule biologics, especially using high-resolution accurate mass spectrometry. Currently, she is a Senior Scientist at Pfizer. She is now working in the department of Pharmacokinetics, Dynamics and Metabolism, focusing on the bioanalysis and ADME profiling of new biologics entities, such as antibody, antibody-drug conjugate (ADC) and fusion proteins.
Abstract:Profiling ADME attributes of Antibody-Drug Conjugates Antibody-drug conjugate (ADC) is designed for preferential delivery of active pharmacological agent to the cellular target. Measuring conjugated drug of the ADCs is a desirable approach for better assessment of exposure, at the mean time the conjugation of cytotoxic agent to the antibody poses a challenge for precise quantitation of the active moieties. Conventional ligand binding assay is highly dependent on the competency of the anti-payload reagents and may not always capture the heterogeneous nature of the ADC, especially when there are active metabolites of the payload. We have established a novel LC/MS strategy for measuring PK of conjugated payload. Depend on the chemistry of linker-payload and conjugation, intact protein analysis, enzymatic cleavage or bottom-up approaches are employed for characterization and quantitative measurement of conjugated active moieties. For cleavable linker based ADC, an enzymatic released protocol was applied to yield the free payload for LC/MS/MS quantitation. For non-cleavable linker conjugation, drug loading profile can be analyzed by intact protein LC/MS method. An alternative approach is to identify the signature peptide which contain the conjugation after enzymatic digestion, and followed by LC/MS/MS quantitation. This conjugated payload strategy has being utilized for the development of novel linker-payloads. Overall ADME evaluation of ADC candidates requires multiple platform approach. We will present some of the cases where in vitro assays can be employed to extrapolate efficacy and toxicity profile in preclinical models.
Mary Joan Castillo
Mary Joan Castillo obtained her Bachelor’s degree at the University of the Philippines in 2007 and immediately started career in the pharmaceutical industry and worked for the R&D Analytical Chemistry Division at United Laboratories (UNILAB) in Manila, Philippines, where she developed and validated assays for the analysis of active pharmaceutical ingredients in new formulations. After almost two years, she then went to work at the University of Santo Tomas Center for Drug Research, Evaluation, and Studies (CeDRES) and performed bioanalytical analysis of pharmaceuticals and metabolites in blood for bioequivalence clinical trials for more than a year. In 2010 she went to start Ph.D. program at the University of Connecticut in Xudong Yao lab, with her research on MS-based proteomics studies, mainly for the development of high throughput methodology for biomarker validation applications. While in graduate school, she went for internship in the Bioanalytical Sciences Division of Biopharm R&D in GlaxoSmithKline (Pennsylvania for 6 months). She will be starting in Synlogic as Associate Scientist in December.
Abstract:Targeted quantitative proteomics for cancer biomarker validation via high sample throughput N-in-1 uMS; Mary Joan Castillo, Adam J. McShane, Min Cai, Yuanyuan Shen, Lei Wang, and Xudong Yao. Recent advances in mass spectrometry (MS) instrumentation and methodologies present rapid and reliable discovery of disease-relevant proteins as biomarkers. Their application in the validation phase, however, is limited by their sample throughput. We leveraged the intrinsic analyte multiplexing capability of MS to sample multiplexing that breaks through the sample-throughput bottleneck of current MS methods. We termed the new methodology as ultrathroughput MS (uMS), specifically ultrathroughput multiple reaction monitoring MS (uMRM MS), and demonstrated its application through one-experiment quantitation of multiple serum samples containing common, low-abundance prostate specific antigen (PSA). The novel bioanalytical platform integrates non-isotopic peptide derivatization, single peptide-level enrichment technique, and MRM MS analysis to perform N-in-1 quantitation of signature peptides from low-abundant proteins. Short peptides with varying sequences and N-terminal capping groups (termed as peptidyl reagents) were prepared for uMS high signal yield screening. The open-source design of uMRM MS technology offers facile adaptability and unprecedented potential for improving the sample throughput in targeted proteomics quantitation using economical, signal enhancing, and structurally customizable reagents to facilitate MS-based validation of cancer protein biomarkers.
Questions? Need to contact the organizers?