Short Courses

Monday, October 18

Principles of High-Throughput Screening for Drug Discovery

High-throughput screening (HTS) is an essential component of modern drug discovery encompassing many domains: biology, analytical chemistry, hardware automation and robotics, software automation, workflow optimization, and quality assurance, just to name a few.   In this course, we will deconstruct HTS for purposes of elucidating its many domains, while simultaneously maintaining a practical emphasis.   In addition to discussing collection of data, we will also examine strategies for efficient conversion of data into information using a variety of tools, such as spreadsheets (e.g. Excel), programming languages (e.g. Visual Basic), relational databases (e.g. Microsoft Access), and advanced visualization tools (e.g. Spotfire DecisionSite).   Case studies from ADME and combinatorial screening will be presented, with mass spectrometry as the key analytical technology.

Instructor: Eric Milgram, Advanced Chemistry Development

Peptide Sequencing and Protein Identification by µLC/MS/MS Ion Trap Mass Spectrometry

This course is a distillation of experiences related to the use of ion trap mass spectrometers for the low level sequence analyses of peptides and proteins. It covers both fundamental principles and useful practices that will facilitate the use and application of µLC/MS/MS. Easy-to-follow mini-labs give attendees hands-on experience in µ-column construction and nano-spray source optimization. The analytical benefits of nanospray will be discussed, and keys to success for robust µLC and nanospray will be emphasized. Example applications direct participants through a step-by-step process of analyzing digested proteins, reviewing the data, and interpreting the results. The target audience includes chemical and biological scientists who want to learn how to begin identifying peptides and proteins using µLC/MS/MS ion-trap mass spectrometry. Some MS experience is recommended because basic LCMS and LCMSMS techniques are not directly covered. Examples will be based on the Thermo LC, LTQ, LTQ-FT ion traps.

Outline:

• Ion Trap Mass Spectrometry: key concepts of operation
• Moving from microspray to nanospray on commercial instruments
• Analytical benefits of nanospray and µLC
• Micro-capillary HPLC: pumps, autosamplers, and column construction techniques (mini-lab)
• Nanospray source tuning (mini-lab)
• Performing m LC/MS/MS on the ThermoFinnigan LCQ (step-by-step example)
• Interpretation of LCMSMS data and sequence elucidation methods
• Making use of advanced instrumentation: LTQ and LTQ-FT

Instructors: Nathan Yates, Merck Research Laboratories & Gary Valaskovic, New Objective, Inc.

Metabolic Profiling: Principles and Practice

The profiles of biological fluids contain a vast array of endogenous low-molecular weight metabolites, the composition of which depends upon the sample type (plasma urine, bile etc) and factors such as the species, age, sex, diet of the organism from which the sample derives and indeed even the time of day at which the sample was taken. Disease, drugs (and other biologically active molecules) perturb concentrations and fluxes in intermediary metabolic pathways. The response to this perturbation involves adjustment of intracellular and extracellular environments in order to maintain homeostasis. Both perturbation and adjustment are expressed as changes in the normal patterns in biofluids or tissues that are characteristic (a 'fingerprint') of the nature or site of the disease process, toxic insult, pharmacological response or genetic modification. These patterns can be evaluated using spectroscopic techniques combined with multivariate statistical methods to obtain insight into the response of a biological system to these perturbations in a time-related manner. To understand and interpret the data from these metabolic profiles it is vitally important to identify and characterize the metabolites both known and unknown when they are observed. This intent of this workshop is to describe methods for conducting metabolic profiling studies and to illustrate how the metabolites of both xenobiotic and various structurally endogenous metabolites can be identified using modern spectroscopic techniques either alone or by employing hyphenated methods.

Instructors: John Shockcor, Bruker BioSpin; Gabi Zurek, Bruker Daltonics;
Andrew Nicholls, GlaxoSmithKlin

Thursday, October 21

Sample Preparation and HPLC Method Development for LC/MS

This practical course on sample preparation and HPLC method development for LC/MS covers the specifics of how sample preparation and HPLC methods are developed for interfacing to ESI and APCI. The course discusses the importance of sample preparation and HPLC when interfaced with mass spectrometry including the extraction and HPLC separation of undetected matrix components, isomers and labile metabolites. The development of sample preparation and HPLC methods for LC/MS for use in qualitative and quantitative analysis throughout drug discovery and drug development are presented. Several types of sample preparation strategies are discussed including SPE (offline and online) column switching (low and high flow applications), Liquid-Liquid extraction and Protein Precipitation. The most advantageous column chemistry for each LC/MS application are thoroughly discussed including, reverse phase, normal phase, ion-exchange, HILIC, monolithic and polar embedded phases. Step-wise method development tutoria ls of developing methods for specific compound classes based on the structure are included in the final summary for both HPLC and sample preparation.

Instructors: Shane Needham, Alturas Analytics & Emile Koster, Spark Holland