CPSA USA 2015

Short Course 1 - Princeton 1 Ballroom
Protein ID and Biomarker Analysis by Mass Spectrometry: Challenges and Solutions for Quantification and Differential Analysis

Mass spectrometry, as enabled by modern ionization methods such as MALDI and ESI, has dramatically transformed the science of protein identification and quantification. There is a rapidly growing demand for the quantitative analysis of proteins and peptides by LC-MS/MS in the bio-analytical and clinical laboratories. This short course will help attendees deeply appreciate specific challenges associated with successful outcomes in LC-MS based protein analysis. An in-depth view of the practical aspects involved in protein/peptide analysis by LC-MS/MS will be covered and will include case studies from the literature and from the instructor's personal experiences.

The use of nanospray enabled LC-MS has developed from a qualitative tool for (global) proteomics to a quantitative method suitable for activities such as peptide/protein biomarker validation. Key to success has been the combination of highly specific sample preparation methods, high sensitivity nanospray ionization, and high performance tandem mass spectrometry. Critical parameters involved in robust sample preparation, nanobore LC and nanospray, sensitive & selective MS detection along and their analytical benefits will be emphasized. The transition of the qualitative nLC-MS/MS technology to methods suitable for absolute quantification will be discussed. New tools for cloud based data sharing and the underlying principles for biomarker identification and validation by differential mass spectrometry will be presented. Real world examples will be presented. The course will also feature an additional hands-on "connections workshop" to hone your expertise is making high quality LC-MS fluid connections.

The target audience includes analysts that have been engaged in qualitative proteomics wishing to transition to quantitative methods, analysts that have specialized in small molecule quantification, and those involved in the development of methods for clinical outcomes.

Instructors:
Nalini Sadagopan, Agilent Technologies
Gary Valaskovic, New Objective
Nathan Yates, University of Pittsburgh

Short Course 2 - Princeton II Ballroom
New! How to Teach an Old Dog New Tricks; The Use of Traditional and Novel Methods, Practices, Technologies to Quantify Protein Therapeutics in a Regulated Environment

This course will give an overview of the methods, practices, technologies and instrumentation used to perform LC-MS in the regulated bioanalytical laboratory. Discussions will center on how fundamental sample preparation techniques such as precipitation, SPE, SLE and LLE, can be utilized to quantitate proteins, peptides, and biomarkers. The process of “bottom-up” digestion based quantitation will be discussed in detail. In addition to traditional sample preparation techniques, the course will present how to use of Immunoaffinity Purification to improve sensitivity and selectivity. Specific examples will be used to demonstrate how to avoid the pitfalls during method development/validation. This course will feature a dynamic format where the latest technology and the advantages of the latest LC-MS instruments will be highlighted. A portion of the course will be devoted to, "validating LC-MS instruments for "21 CFR Part 11" compliance. At the end of the course, a method development tutorial will be presented to the participants..

Instructors:
Shane Needham, Alturas Analytics
Jim Shen, Bristol-Myers Squibb
Dave Abramowitz, AB SCIEX

Short Course 3 - TBA
Human Pharmacokinetic Prediction and Approaches to Identify Rate Determining Step of Drug Absorption and Elimination: Industrial Perspective

Human Pharmacokinetic Prediction and Approaches to Identify Rate Determining Step of Drug Absorption and Elimination: Industrial Perspective The oral route is the predominant way of delivering drugs to the systemic circulation to generate pharmacological effects and clinical benefits. To ensure steady flow of new drugs to patients, the pharmaceutical industry is focused on designing and developing molecules to achieve systemic drug exposures capable of robustly modulating disease relevant biological mechanisms in humans. Solubility, membrane passive permeation, transporter mediated active efflux or influx and metabolism are the main factors contributing to oral absorption, disposition and elimination. Therefore understanding the interplays of these factors is crucial to overcome the challenges of poor bioavailability and pharmacokinetics and regulatory concerns of drug-drug interactions in GI absorption. On the other hand, clearance rate is a critical determinant of drug exposure in the systemic circulation and consequently at the pharmacological target compartment. Hence, it is a key component in determining the therapeutic efficacious dose. Either drug metabolic stability or active hepatic uptake could be a rate determining process of drug clearance. Identifying the rate determining step in drug clearance early on in drug discovery is a prerequisite for successful drug development. Furthermore, Drug-Drug Interaction (DDI) is one of the leading causes of morbidity in United States. This is of particular concern in patient populations that are treated with multiple different types of medications (i.e. polypharmacy), such as those suffering from cardiovascular, metabolic and endocrine diseases (CVMED). This practical/hands on course is specifically designed for personnel in the pharmaceutical and biotechnology industries and contract research organizations (CROs), who need to understand:

• General principles of pharmacokinetics
• Factors that affect drug absorption with emphasis on the impact of intestinal transporters on drug absorption
• Identification of rate determinant step of drug clearance and introduction to extended clearance classification system (ECCS) and its application in drug discovery
• Review of major metabolizing enzymes and in vitro/in vivo tools to predict human metabolism mediated clearance
• Review of known renal/hepatic transporters and their role in drug disposition and elimination
• Review of principles of transporter mediated drug interactions and static models used to predict them
• Overview of FDA guidance on assessment of transporter mediated drug interactions
• The workshop will also include a hands-on session that aims at improving your ability to apply these strategies to medicinal chemistry for hit selection, lead optimization, and development candidate selection

Instructor:
Ayman El-Kattan, Pfizer
Yurong Lai, Bristol-Myers Squibb

Short Course 4 - Lehigh
Building a Better Clinical Mass Spectrometry Lab

Clinical Diagnostics is growing in the use of mass spectrometry and especially in the implementation of new techniques. With the growth of pain management, the use of LC-MS/MS has seen the greatest growth. Now that LC-MS/MS is firmly positioned in clinical diagnostics, there exists a need for education in many new ways such as the need to improve education of lab technicians in the use of mass spectrometry and the need to use and expand new technologies to clinical diagnostics. This workshop will provide a unique opportunity to interact with though leaders from both clinical diagnostics and mass spectrometry technologies. Topics will range from the use of new interfaces to the MS to enable ease of use, the development of tests that are easy to implement, and the opportunities available for individuals looking to enter the clinical market place.

Instructors:
Timothy Garrett, University of Florida
Mark Hayward, ITSP Solutions, Inc.
Kenneth Lewis, Opans