About this course
This course is aimed at anyone who wants to develop an understanding of theory and techniques in protein purification using fast protein liquid chromatography (FPLC) on an ÄKTA chromatography system. The course, while specifically designed around the ÄKTA Avant system, provides comprehensive insights and practical knowledge that will be beneficial to all FPLC users, regardless of the specific model they are using. This training course will also focus on maintenance, cleaning and applications of using FPLC.
The 3-day course consists of lectures, practical demos and hands-on exercises, hosted by the Industrial Biotechnology Innovation Centre (IBioIC). On completion of the course, the attendees will be presented with a certificate of course completion by IBioIC.
Learning outcomes
Learn the basic principles and applications of different chromatography techniques, such as affinity, ion exchange, hydrophobic interaction, mixed mode and size exclusion
Learn how a chromatography system works, how to prepare for a chromatography run, and importantly, how to ensure your chromatography system remains clean
Purify green fluorescent protein (GFP) using Ion exchange chromatography (IEX) and hydrophobic interaction chromatography (HIC). Qualitative and quantitative analysis of purified protein using SDS-PAGE and UV-Vis spectroscopy
Learn how to pack chromatography columns like a pro through a live demonstration and discover why it matters for your chromatography results
Who will benefit?
Scientists, engineers, graduates, technical managers and operators, especially those in their early careers in downstream bioprocessing
Anyone who would like to start using a chromatography system or wants to gain an understanding of the theory behind chromatography techniques
Please contact scaleup@ibioic.com for further details and sign-up
Course schedule
Day 1
Lecture 1: (9:15-10:00)
Introduction to protein purification
Prayag Poreri, Senior Downstream Processing Scientist, IBioIC
Lecture 2: (10:15-11:00)
Virtual Reality in Bioprocessing: A Hands-On Training Experience
Laura Porcza and Nicole Barnes, Resilience
Practical 1: (10:15-12:00)
Introduction to ӒKTA Avant chromatography system: Familiarising with system pumps, system and sample inlets, sample pump, mixer, UV, pH, conductivity monitor flow restrictor and fraction collector
Practical 2: (13:00-16:00)
Introduction to UNICORN™ software:
Familiarising with Administration, System control, Method Editor and Evaluation tabs
Manual operation of ӒKTA Avant,
Method creation (basic and advanced)
Day 2
Invited Lecture 2: (9:15-10:00)
Antibody purification using FPLC (TBD)
Lecture 3: (10:15-11:00)
Protein characterisation tools
Prayag Poreri, Senior Downstream Processing Scientist, IBioIC
Practical 3: (11:15-12:00)
Packing and equilibration of a chromatography column: initial manual packing, connecting the column to ӒKTA Avant and final system-assisted packing.
Equilibration of the packed column
Practical 4: (13:00-14:30)
Protein capture: IEX Purification of green fluorescent protein (GFP) using a manual run
Evaluation and peak analysis
Column maintenance and storage
Practical 5: (14:45-16:00)
SDS-PAGE analysis of IEX-purified fractions, fraction pooling and storage
Day 3
Invited Lecture 4: (9:15-10:00)
Choosing the right resin and Scaling up (TBD)
Practical 6: (11:15-12:00)
Final polishing: further purification of the IEX-purified sample using a HIC chromatography method run. Evaluation and peak integration,
Column maintenance and storage
Practical 7: (13:00-14:30)
SDS-PAGE analysis of HIC-purified fractions, fraction pooling, UV-Vis spectroscopy analysis and storage
Practical 8: (14:45-16:00)
Maintenance of ӒKTA Avant: pH calibration, system and pre-column pressure calibration, changing pump rinsing solution and mixer filter, final CIP and storage of ӒKTA Avant.