Practical HPLC for Biopharmaceuticals

Custom Content

To find out more about customizing this course to meet your specific needs, give us a call at
(925) 297-5374

or email us at info@lcresources.com

Virtual On-Site: 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 5 - 30 people using Citrix GoToMeeting service. 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