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Printable Handouts
Navigable Slide Index
- Introduction
- Separation processes that can be used to fractionate proteins
- Chromatography: historical perspective
- Types of column chromatography
- Ion exchange chromatography: general principles (1)
- Ion exchange chromatography: general principles (2)
- Ion exchange groups
- Size-exclusion chromatography: gel filtration
- Types of affinity chromatography
- Tags to facilitate affinity purification of recombinant proteins
- Chromatography components (1)
- Chromatography components (2)
- Chromatography equipment (1)
- Chromatography equipment (2)
- HPLC of proteins
- Summary of recent chromatography improvements
- Recent laboratory scale (analytical becomes preparative)
- Troubleshooting chromatography problems (1)
- Troubleshooting chromatography problems (2)
- Scaling up protein purification
- Challenges of protein purification for pharmaceutical applications
- References
- Conclusions
Topics Covered
- History of chromatography
- Forces and properties of proteins as handles for fractionation
- Types of chromatography
- Chromatography components
- HPLC
- Trouble shooting some chromatography problems
- Protein purification for pharmaceutical applications
Talk Citation
Burgess, R.R. (2020, July 30). Fundamentals of protein chromatography: protein chromatography [Video file]. In The Biomedical & Life Sciences Collection, Henry Stewart Talks. Retrieved October 11, 2024, from https://doi.org/10.69645/JWTV3730.Export Citation (RIS)
Publication History
Financial Disclosures
- There are no commercial/financial matters to disclose.
Fundamentals of protein chromatography: protein chromatography
Published on July 30, 2020
46 min
A selection of talks on Methods
Transcript
Please wait while the transcript is being prepared...
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.