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The workshops will mirror important developments in
MicroScale (Bio)separations.
They intend to provide an entry into the topics of the symposium to novices in to the
field, to PhD and post-doctoral students and/or to experienced users who want to
obtain a comprehensive view of the particular topic. The workshops will last half a
day and will be arranged such that one can participate in two coherent workshops.
- Capillary Electrophoresis Method Development – Prediction and Optimization of Separation
Bohuslav Gas, Charles University of Prague, Department of Physical
and Macromolecular Chemistry
The workshop will help participants to understand electrophoresis and electro-migration
in general. It intends to provide an entry into the topic both to newcomers to
understand basic principles and experienced users who want to improve separation
in systems of interest. Using computer simulation tools, PeakMaster and Simul, (see
http://www.natur.cuni.cz/gas) the participants are able to simulate real experimental
runs and inspect behavior of separation systems, regardless whether in classical capillaries
or in chips. Both computer programs run in fully graphical environment and
enable: (i) to calculate pH of background electrolytes, (ii) to predict a possible occurrence
of system peaks and find the ways to avoid them, (iii) to adjust composition
of the background electrolytes for getting maximum resolution, maximum detector
signal and minimum electro-migration dispersion (minimum triangulation).
The workshop brings a chance for participants to perform their own “virtual separations”
in particular systems and inspect the suitability of the electrolytes used. The
composition of the background electrolytes can be then optimized to reach better
separation without a necessity to use “wet chemistry”. In the method development
this can spare a considerable amount of experimental time and chemicals.

Miniaturization has become a key component in the concept of comprehensive genomic
and proteomic analysis, mainly triggered by the availability of very low amounts
of sample in biological analysis. In this workshop we therefore focus on single- and
multidimensional separations of biomolecules carried out in miniaturized analytical
systems, employing capillary devices or microchips as separation medium. An overview
of the different separation modes is given, followed by information on the state
of the art in instrumentation and column technology developed to operate the
single and
multidimensional analytical systems. Subsequently, on-line or off-line methods
for interfacing of the separations to mass spectrometry are presented. We discuss
the new trends of ultrahigh- pressure chromatography, monolithic separation media,
and chip-based separation devices, which have facilitated a significant increase in
separation performance and speed of analysis. Finally, some selected examples of
application are presented in order to shed light on the potential and the limitations of
micro- and nanoscale separations in genomic and in proteomic analysis.

The aim of the workshop is to give insight in the relevant techniques of metabolomics
and to discuss the analytical strategies and developments for specific applications of
metabolomics. Metabolomics involves the non-targeted analysis of all the metabolites
at sub-cellular, cellular, tissue or body fluid level during changes of a biological system.
In addition, metabolomics is an important part of the Systems Biology concept, i.e. the
study of a biological system as an integrated system of genetic, protein, metabolite,
cellular, and pathway events that are in flux and interdependent. In this course some
state-of-the-art analytical techniques (GC-MS, LC-MS, CE-MS and NMR) will be discussed.
Specific attention will be directed to data pre-processing and data analysis
and knowledge extraction. In addition, metabolite identification will be discussed. Applications
of metabolomics in various biological areas will be used to illustrate the possibilities
and current challenges. Trends and future perspectives will be discussed.

The workshop will compromise of a series of short presentations by
leading industrial experts on their experiences encountered during the
routine application of CE methods in the pharmaceutical /
biopharmaceutical industry.
The presentations will highlight uses of CE in various areas of product
development e.g. product characterization, QC lot release and stability
testing, all areas requiring robust analytical methods.
Examples for efficient method development, method validation and QC
batch release (e.g. for small molecule pharmaceuticals as well as
biopharmaceuticals) are given. Validation parameters (including precision,
accuracy, linearity), acceptance criteria and results of CE assays used in
QC batch release testing of (bio)pharmaceutical compounds will be
discussed. Experiences of method transfer, between analysts, laboratories
and sites will be presented and described. Instrument qualification (IQ/OQ/PQ)
strategies are discussed. The remainder of the workshop will be open for
all attendees to raise questions for the expert panel. It will allow for
open discussions of specific techniques and/or applications. In addition
attendees will be encouraged to share learning’s from their experiences
in routine method operation, method validation and transfer.
Contributors to the workshop:
Cari E. Sänger - van de Griend, Pharmaceutical and Analytical
R&D; AstraZeneca; Södertalje; Sweden
Steffen Kiessig, Protein Analytics and CE - Development; Solvias
AG; Basle, Switzerland
Oliver Grosche, CHAD, Novartis Pharma AG, Basle,
Switzerland
Ilias Jimidar, Analytical Development, Johnson & Johnson
Pharmaceutical Research and Development, Beerse, Belgium
Oscar Salas-Solano, Genentech Inc., San Francisco, CA, USA
Micro-scale separation methods have played an important role in the
development of LC-MS from the early days in the 1970s. Nowadays, the
application area seems to be divided into two camps: high-throughput
analysis at high flow-rates for among other bio-analysis, environmental
monitoring, food safety analysis, and low flow-rate applications in
biochemistry, proteomics and related areas. This workshop focuses at the
principles and applications of micro-scale separation in on-line
combination with mass spectrometry. The discussion will be primarily
technology-oriented: how does it work, and what to choose for a particular
application. After a brief introduction in the various ionization
techniques (electro-spray, atmospheric-pressure chemical and
photo-ionization, and matrix-assisted laser desorption/ionization) and the
ways to couple these to various micro-scale separation methods, the
attention is focused to the principles and merits of the various types of mass analyzers
available, i.e., (triple) quadrupole, (linear) ion-trap, time-of-flight,
sector-field, Fourier-transform ion-cyclotron resonance instruments, and
combinations thereof. The fundamental discussions are enlivened and
illustrated with some typical applications of various approaches. The
approach is pragmatic and practice-oriented rather than strictly
theoretical. There is ample time for discussion and questions
.

Microfabricated fluidic devices have received steadily increasing attention from both
the research and user communities over the past several years. These devices hold
much promise for addressing a number of chemical and biochemical analysis problems
in areas ranging from the biotechnology and pharmaceuticals industries to environmental
monitoring. This workshop will be tutorial in nature and attempt to familiarize
attendees with developments in this field of research that have taken place over
the past decade. A cursory discussion of fabrication principles used to make these
devices will be covered in addition to basic microfluidic operational principles. The
focus of the workshop will be on the use of micro-fabricated fluidic devices for solving
chemical separations problems. The advantages and disadvantages of performing
separations in miniaturized formats will be reviewed. A brief overview of commercially
available microfluidic devices will be given as well as a discussion of development in
the research community. We will also cover the integration of functional capabilities
such as chemical reactions and sample processing with chemical separations.

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