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Immuno-assay for screening

screening cytokines immuno-assay compounds

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#1 amdspieg

amdspieg

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Posted 08 September 2012 - 11:13 AM

Hey,

I'm a medical student and for my research project I've got to screen two hundreds of new compounds for their possible immuno-activating role (a new group of compounds on which no data in literature, hypothesis is that they may act on receptors/proteins of immune-cells).

Because it's a screening, I'm looking for a general immune test, where sensitivity is much more important than specificity (I don't want false-negatives). In later stages we can do more refined test to determine on which cells they act, how they act,...

Some suggestions:

1) splenocyte-culture with/without stimuli + compound=> measuring cytokines with elisa (IL2, IFNg, IL4, IL6, TNFa); the same with hPBMC (more monocytes); isolated human neutrophils tests with respiratory burst/degranulation test/migration test/Ca test

2) is it possible to have the neutrophils and PBMC in one sensitive assay? eg. measuring cytokines in whole blood (il-8 for neutrophils)? Will I not miss neutrophil-activating compounds that do not enhance IL-8 production (but do enhance respiratory burst,...)?

Other suggestions? Thanks in advance!





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