Hi,
I am working on a project which involves looking at T cell responses to
recombinant bacterial proteins. One problem I have been having is that
some of the recombinant proteins that I have been expressing and affinity
purifying totally crash out of solution when I dialize out the denaturant
(8 M urea). I have tried a number of standard tricks to try to keep these
proteins in solution with no avail including; 1) Step dialysis into an
intermediate urea concentration solution, 2) Dialysis into 10 mM Tris (pH
8.0). 3) Dilution of protein prior to dialysis. So, in regards to this
problem I have a few questions; 1) Does anyone have any ideas of
additional things to try in order to keep my proteins in solution?, 2)
Are there any detergents that I can use that might be compatible with T
cell proliferation assay (mixed lymphocyte assay using rabbit
splenocytes). 3) Are T cell responses to insoluble particulate antigens
comparable to that of soluble antigens. As per my last question I have
read that macrophages will take up, process and present particulate
antigens more efficiently than soluble antigens, is this generally true?
Thanks,
Thomas
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Thomas W. Arroll
Department of Pathobiology
University of Washington
Seattle, WA 98195
(206)731-8016
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