Fundamentals of protein chromatography: protein chromatography

Published on July 30, 2020   46 min

A selection of talks on Methods

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0:00
Hello, I'm Richard Burgess. I'm a Professor Emeritus of Oncology at the University of Wisconsin in Madison. My title of my talk today is fundamentals of protein chromatography part 2. Part 1 was about the basics of protein purification and part 2 will be focused more on protein chromatography.
0:22
This slide shows a repeat of one from part 1, which is basically different separation processes that can be used to fractionate proteins. Again, there are a variety of processes, precipitation, chromatography, electrophoresis, etc. What we're going to focus on today is the chromatography part; the various kinds of chromatography that can be used to fractionate a mixture of proteins in order to work toward the purification of a particular protein.
0:53
Before I get into the meat of my talk, I wanted to just give a little bit of historical perspective. Chromatography hasn't been around forever. It was really embedded in 1903 by a Russian scientist, Tswett, who was a botanist who was separating plant pigments and ran a mixture of pigments through a solid column of resin and noticed that the pigments separated into different colors which could then be eluted from the column and studied. This is really where the term chromatography came from. The pigments were colored. Most of us use the term chromatography, but work with solutions which are not colored. Anyway, this was something that was done well over 100 years ago and has developed into a very powerful fractionation, not only for pigments in small molecules, but for larger molecules such as proteins. In general, the chromatography is done on some substrates like paper chromatography, thin layer chromatography, and then low-pressure column chromatography, and then more recently, high-pressure or high-performance liquid chromatography. There's been a progression to more and more powerful in high resolution methods.

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