Immunoaffinity chromatography and gentle IAC purification of complexes

Published on May 30, 2021   36 min

Other Talks in the Category: Biochemistry

0:00
I'm Richard Burgess, I'm James D.Watson Professor Emeritus of Oncology in the McArdle Laboratory for Cancer Research at the University of Wisconsin-Madison. Today I'd like to talk about the powerful protein purification method, immunoaffinity chromatography, and in particular how we've developed some very gentle immunoaffinity chromatography purifications that allow us to isolate labile complexes.
0:31
The first use of the immunoaffinity chromatography was in the early 50s. It wasn't what I call immunoaffinity chromatography, it was actually putting an antigen (the thing that the antibody binds to), attaching it to a cellulose column and then using it to purify an antibody. As I think of it, immunoaffinity chromatography is where you usually attach an antibody to the resin, and use it to purify a protein containing an antigen. In 1968, Pedro Cuatrecasas published a very important paper which introduced the concept of affinity chromatography, where you can attach something to a chromatography resin and use that to pull out a protein that binds to that affinity material, whether it be a small molecule or an antibody or anything else. This was an important development, and there are many different kinds of affinity chromatography now being used. The other really important development was in 1975, when Kohler and Milstein published their paper on the ways of purifying what are called 'monoclonal antibodies'. In this case, isolation of a B-cell hybridoma which produces a single antibody to the antigen of interest. Because this is a cell line that is from a single clone, it's called a 'monoclonal antibody', and this has turned out to be extraordinarily important for high-specificity immunoaffinity chromatography, and many other things in the area of therapeutics and diagnostics.
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Immunoaffinity chromatography and gentle IAC purification of complexes

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