Course Description
In this course you will learn how to develop methods and implement CE for the separation of biomolecules. The course begins with an extensive review describing the factors that influence the electrophoretic migration of molecules in fluid solution. A phenomenological approach is employed that enables the student to take home a thorough understanding of electrophoretic processes. The curriculum includes:
Modules covering the modes of capillary electrophoresis
- Capillary zone electrophoresis including micellar electrokinetic separations
- Capillary isoelectric focusing
- Capillary sieving electrophoresis (size separations)
Methods development using each of the CE modes
Applications including amino acids, peptides and proteins, SDS-proteins and carbohydrates
DNA applications including oligonucleotides, sequencing, plasmids, short-tandem repeats and genetic analysis
In addition, you will learn:
- How to perform quantitative analysis
- How to develop robust methods
- Sample preparation methods for CE
- Why dynamic coatings are important
- The advantages and disadvantages of each CE injection technique
- Various detection techniques such as UV, LIF and mass spectrometry
- Why on-capillary detection is different from post-column detection
- How to logically troubleshoot a malfunctioning method
- How to implement stacking procedures to overcome sensitivity problems
- How to perform method validation, methods transfer and failure analysis
- The types and availability of commercial equipment
- Why microfabricated systems represent the next generation of CE instruments
Complete Course Agenda
Introduction to Capillary Electrophoresis
- The transition from slab-gels to capillaries
- Using diagrams to help understand the impact of ion migration on separations
- How to optimize resolution
- Buffers and buffer additives
- Commercial instrumentation
Capillary Zone Electrophoresis
- Effective use of mobility plots
- How to control wall effects
- How to develop a well controlled separation
- When to use and how to optimize secondary equilibrium and micellar electrokinetic separations
Capillary Isoelectric Focusing
- Introduction to isoelectric focusing
- How to select instruments, capillaries and ampholytes
- Case study, erythropoietin
Capillary Sieving Electrophoresis
- Introduction to polymer networks
- Method development strategies
- Size separations for proteins and DNA
- Case study, monoclonal antibodies
Injection
- When to use and how to optimize both electrokinetic and hydrodynamic injection
Detection
- Why corrected peak areas are required with CE
- Types of detection including mass spectrometry (MS)
- Case study, carbohydrate separations with detection by laser induced fluorescence and MS
Troubleshooting Guide
- Common and unusual problems are described
Stacking and Trace Enrichment
- How to optimize stacking to solve sensitivity problems
Microfabricated Systems
- Why microfabricated systems represent the next generation of CE
Validation and Methods Transfer
- Experiences at large biotech companies
- Case studies
- Failure analysis
Course Instructor
Dr. Robert Weinberger, Ph.D., President, CE Technologies, Inc.
Dr. Robert Weinberger is a consultant specializing in the field of Capillary Electrophoresis (CE) and related technologies. Formally of Applied Biosystems Inc., he has been involved in CE since 1987. He is currently President of CE Technologies, Inc., a consulting firm specializing in contract research using CE. He is the instructor of the American Chemical Society short course entitled "Practical Capillary Electrophoresis" and is the author of the textbook of the same name, now in its second edition. Another short course "Capillary Electrophoresis for Biomolecules" was introduced in 2006. His column on CE appears often in American Laboratory and he is the author of 50 technical publications, patents and book chapters as well as close to 100 magazine articles and meeting reviews. He has presented numerous papers and seminars at scientific conferences, universities and discussion groups in the US, Europe and Asia. Dr. Weinberger is a reviewer for many major journals and sits on several editorial boards. The Journal of Capillary Electrophoresis honored him as its featured scientist in 1999. He served as an expert witness in CE patent litigation surrounding DNA sequencing.
