Antibodies are one of the most common products made by biotech companies. They can be used to block viral infections and treat many forms of cancer. But first, we need to grow the cells that make them (biomanufacturing - upstream processing) and then, we need to purify them (downstream processing).
In this week's Friday webinar, Dr. Margaret Bryans will tell us about an on-line module from Montgomery County Community College for biomanufacturing where students learn about downstream processing and antibody purification.
Bruce Van Dyke from Quincy College will show us how to use a really neat online site from the late Andrew Booth where students can explore the use of different kinds of chromatography for purifying proteins and designing purification strategies.
How do you teach concepts in protein purification without a lab?
Protein Purification is an award-winning program which has been widely used in schools, colleges and universities since 1983. It guides students through some of the more commonly-used protein separation techniques and lets them experiment. It starts off by letting students examine how a simple mixture of proteins behaves during gel filtration and ion-exchange chromatography and then goes on to allow the design and testing of full purification protocols using more complex mixtures of proteins.
The program assumes that users are familiar with the theoretical background to the most common separation techniques, enzyme assays etc. and that they understand the concept of the isoelectric point of proteins.
Here is an example of what we'll do and some results I obtained from a quick purification trial with the iPad app.
The program is very complete. You test your results with Ammonium sulfate fractionation, heat treatment, gel filtration, ion exchange chromatography, hydrophobic interaction chromatography, and affinity chromatography. This would be a great companion to Chromatography techniques Course in Box.
My goal was to try and purify protein #2 from a mixture of three proteins. I started by choosing a DEAE-cellulose column and picking the salt gradient. Proteins that have a negative charge at pH 7.0 will bind to the column and others will flow through.
I "ran" a gel to see confirm the location of my protein.
Last, I did an immunoblot (aka Western blot) to confirm that my protein was in fraction 20, and not yet pure.