Detergent compatability in T cell assays?

Guy Hermans ghermans at
Tue Oct 28 06:41:02 EST 1997

In article
<Pine.OSF.3.96.971027112503.9217A-100000 at>, Thomas
Arroll <twa at> wrote:

> 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?   

Try dissolving the proteins in pure ethanol (pro analysi quality; filter
sterile after dissolving the protein). Sometimes smaller quantities (say,
10%) will to.

Second solution (pun intended) is using a low (0,01%) solution of Tween-20.

Another option would be to use a 10%DMSO solution - but don't try this one
untill all else fails. DMSO is quite toxic. 

It's absolutely imperative to filter the solution after dissolving the
solution. Not only for sterility reasons: microparticles might still be
present and mess up protein determination assays afterwards, or may
influence the behaviour of the cells.

Good luck,


Guy Hermans, PhD student
MS research Unit             Tel 0032 (0) 11/26.92.07
Dr. L. Willems-Institute     Fax 0032 (0) 11/26.92.09
University Campus            E-mail ghermans at
B-3590 Diepenbeek

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