ISPPP 2007 Short Course Description and Schedule
Sunday, October 21, 2007
The ISPPP Symposium reserves the right, without notice, to modify or amend the roster of Sunday short courses and/or presenters. Any changes will be updated on the web site.
Short Course # 1: Monolithic Columns: How to Make and Use Them
9:00 AM - noonPresented by Prof. Frantisek Svec, University of California, Berkeley, CA, USA
This workshop will give an introduction to the basics of monolithic stationary phases, their preparation, and selected applications. First, a brief history of monolithic stationary phases, their rebirth at the end of the 1980’s and their fast development ever since will be presented. The various approaches to the stationary phases with reduced discontinuity including aligned fibers, rolled textiles, as well as monolithic discs, and columns based on both silica and synthetic polymers will also be introduced. Then, several specific examples of the preparation of monolithic columns based on synthetic polymers will be shown with emphasis on simplicity of both thermally and UV initiated processes and variety of chemistries easily available. Approaches to larger scale monoliths for preparative separations will be described. Also, monolithic materials placed in the currently very popular capillary and microfluidic formats will be presented in more detail. In addition, methods leading to desired chemistries by grafting of pores with selected functional monomers and combinations of various chemistries and functions within the same monolith will also be described. Due to the specifics of monolithic columns enabling high flow rates without compromising the efficiency, high speed/high throughput separations of a variety of compounds including proteins, peptides, nucleic acids, synthetic polymers, and small molecules in HPLC mode can be achieved. Several examples of these separations using monoliths of very diverse shapes and sizes from very large radial flow columns, over analytical scale units, to capillary columns will be shown. Since monolithic columns also represent quite a significant share of all columns used in capillary electrochromatography, their preparation and use in CEC will also be presented. This workshop will be wrapped up by discussing current trends and future developments of this promising new format of stationary phases.
Short Course # 2: Mass Spectrometry in Glycomics and Glycoproteomcs
9:00 AM - noonPresented by Prof. Ron Orlando, The University of Georgia, Athens, GA, USA
Glycosylation is one of the two most common post-translational modifications found on proteins. Glycan structures and sites of glycosylation have been shown to change with the state/condition of the cell in which the proteins are produced. For example, it has been known for over 40 years that cancerous cells attach different glycans than those of corresponding “normal” cells from the same tissue/organ. Since many glycoproteins are excreted, altered glycosylation has the potential to be used as a biomarker for cancer. Numerous other disease states, ranging from arthritis to alcoholism, are also characterized by altered glycoprotein glycans, as is normal cell growth, differentiation, and development. Identifying glycan structures and how these structures change as cells differentiate or as tumor cells progress, for example, is the focus of an emerging field called glycomics. This workshop will focus on the role of mass spectrometry in the emerging field of glycomics and glycoproteomics. An overview will be presented on the biosynthetic pathway that leads to protein glycosylation and how this, in turn, leads to diverse structures of glycoprotein glycans. Other topics that will be discussed include: the analytical challenges of characterizing glycoproteins and their glycans; the methods used to determine glycan structure, sites of glycosylation, and identify glycans present at individual glycosylation sites. Approaches used for comparative glycomic studies will also be covered. Many of the techniques discussed are applicable to both whole cell glycoprotein extracts (i.e., glycomics) as well as the characterization of purified glycoproteins. Although the emphasis of this workshop will be on N-linked glycosylation, the methodology discussed can be extrapolated to other types of glycosylation
Short Course # 3: Preparative-scale Separation of Biomolecules
1:30-4:30 PMPresented by Prof. Alois Jungbauer, University of Natural Resources and Applied Life Sciences, Vienna, Austria
Chromatographic methods play a pivotal role in biotechnology and biopharmaceutical technology, particularly for high molecular mass compounds such as proteins and plasmids. The high level of purity can be only achieved by chromatographic methods. Beside bulk contaminants traces of bioactive compounds such as endotoxins, DNA and other adventitious agents must be efficiently removed from the process solution. In the workshop special emphasis will be put on the description of the characteristics of chromatography media used in bioseparation and how they differ from analytical media and media used for separation of small molecules. Process optimization, scale up and important design criteria will be discussed. The influence of mobile phase composition on resolution , and guidelines for the optimization of selectivity will be presented. An overview on novel stationary phases for protein and polynucelotide separation and examples for novel bioseparation processes using these phases will be given. The difference and applicability of monoliths, beads with a porous shell and polymer grafted beads will be elaborated. In the second part of the workshop the progress on biorecognition for affinity chromatography will be discussed.
Short Course # 4: Particle Packed Columns and Monolithic Columns in HPLC:A Comparison and Critical Appraisal
1:30-4:30 PMPresented by Prof. Klaus K. Unger and Romas Skudas, Department of Inorganic Chemistry and Analytical Chemistry, Duesbergweg 10 – 14, Johannes Gutenberg-University, 55099 Mainz , Germany
The course is divided into four parts and is thought for beginners in this field as well as for advanced chromatographers. The first part surveys the most important process steps in the manufacture of particle packed columns : synthesis and particle formation, sizing and size analysis, packing procedure and evaluation of column performance.
The second part highlights the most attractive developments in the field of particle packed columns: the ultimate minimum particle size in HPLC – fiction and facts, totally vs. core/shell particles in view of the currently introduced Halo particles, column miniaturization: from meso to micro to nano – where is the end ? and how to gain higher resolution in HPLC ?
The third part discusses the most relevant features of monolithic columns with regard to their structure and chromatographic properties and is divided in three paragraphs: the basic idea and the pioneers, monolithic silica columns, polymer-based monolithic columns. The latter subject will be treated in depth by F. Svec at the short course Number.1 at the beginning.
The fourth part is entitled : comparison of the structure and performance of particle packed and monolithic columns. It summarizes all aspects and will answer the question: Where are we now and where are we going.