Description
On-Site Course: Practical HPLC for Biopharmaceuticals
Who should take this course?
If you are a biochemist or biologist with limited experience using HPLC, this course will acquaint you with instrumental techniques for analysis and lab-scale purification of both biopolymers and small molecules. If you are an analytical chemist with experience in the use of HPLC separations for small molecules, this course will help you apply these techniques to peptides, proteins and nucleic acids.
What does it cover?
Although “small molecule” and “biopolymer” separations have traditionally been considered as separate activities, analysts in the biopharmaceutical industry regularly have to deal with both. Fortunately, the underlying principles of chromatography apply equally well in both situations when interpreted appropriately.
In this course you will learn:
- Fundamentals of HPLC mechanisms of band broadening for both large and small molecules
- An overview of HPLC instrumentation with an emphasis on preventive maintenance and troubleshooting
- Column and mobile phase selection for reversed-phase, ion-exchange and size-exclusion chromatography
- Strategies for developing and optimizing HPLC methods
- HPLC column maintenance and troubleshooting
and much, much more!
You will learn how to chose the appropriate separation technique for your application and how to select the best chromatographic mode to meet your separation goals. You will learn how to develop and optimize an LC method and how to troubleshoot it when problems occur. You will learn how to handle columns to achieve good performance and satisfactory lifetime. The focus of this course will be on practical guides and tips to achieve successful biomolecule separations.
When is the course available?
“The course is presented for groups of 10 – 30 people either “on-site” at your facility or “virtual on-site” using Citrix GoToMeeting service. On-site presentation is two full days. Virtual On-Site is six 2.5-hour sessions (typically 2 or 3 sessions per week). The usual lead time to schedule a course is approximately 4-6 weeks.
Call us at (925) 297-5374 or email to info@lcresources.com to arrange timing, pricing, and content.
What topics are covered?
The course content can be tailored to your specific needs. A typical schedule looks like this:
Section 1. Isocratic HPLC Basics
- What is HPLC?
- Separation Chemistry
- Reversed phase
- HILIC (normal phase)
- Ion exchange
- Ion pair
- Size Exclusion (GFC & GPC)
- Key Measurements: Retention
- Key Measurements: Selectivity, Efficiency, Resolution, & Tailing
- Controlling Resolution
Section 2. Gradient HPLC Basics
- Why use a gradient?
- Gradient is Just Like Isocratic
- Gradient is Different From Isocratic
- Gradient Hardware Issues
Section 3. Reversed-phase HPLC
- Mechanism
- Small Molecules: Solvent Selection
- Small Molecules: Column Selection
- Small Molecules: pH, Additives, & Temperature
- Biopolymers: Solvent & Column Selection
- Biopolymers: Temperature, Denaturation, & Additives
Section 4. Ion-exchange HPLC
- General Mechanism & Packings
- Proteins: Gradients, pH, & Particle Size
- Oligos, Peptides, & Sugars (Oh My!)
Section 5. HILIC (Hydrophilic Intraction Chromatography)
- What is HILIC (and what is it good for)?
- How Does HILIC Work?
Section 6. Size-Exclusion Chromatography (GFC and GPC)
- How Does SEC Work?
- Controlling SEC
Section 7. HIC (Hydrophobic Interaction Chromatography)
Section 8. HPLC Equipment: Reservoirs & Pumps
- Reservoirs and Degassing
- Pumps
Section 9. HPLC Equipment: Tubing & Injectors
- Tubing
- Fittings
- Injectors
Section 10. HPLC Equipment: Columns
- Overview & Physical Problems
- Non-Column Problems (I)
- Non-Column Problems (II)
- Chemical Problems
- Documentation
Section 11. HPLC Equipment: Detectors
- UV Detectors
- Gradient Baseline Issues
- Other Detectors
Section 12. Quantitation
- Measuring Peak Area
- Calibration Issues
- Reproducibility Issues
Section 13. Performance Qualification
- Pump & Detector Checks
- On-line Mixing Checks
- Chromatographic Checks
Section 14. Troubleshooting & Diagnostics
- Preventive Maintenance, Documentation, and Troubleshooting
- A Troubleshooting Checklist
