ELISA coating shelf life question

Dom Spinella dspinella at chugaibio.com
Tue Aug 26 15:02:33 EST 1997


> When running an ELISA is it a problem to coat the ELISA plate with
> antigen and rather than adsorb for ~ 1 hour, leave the plate adsorbing
> long term (several days), if so, will it hurt to do it at 4=B0?
> =

> please respond to my e-mail address: dlarkin at Umich.edu
> =

> thanks for your time
> =

> Dennis Larkin

Dennis:

Ithink it really depends almost exclusively on the antigen in question
and the optimal coating conditions need to be determined empirically. =

For some antigens, overnight (or longer as you propose)coating works
fine; for others a short coat at room temp or 37 degrees works much
better; and for others it makes no difference.  Other variables that can
be optimized include pH and ionic strength of coating buffer,
temperature of incubation, coating matrix (PVC, polystyrene, etc -- and
even different manufacturers plastic formulations vary).  In our hands
these variables can sometimes dramatically influence the ELISA and to
date we have been unable to predict the optimal parameters by limited
knowledge about the physico-chemical protperties of the antigen (e.g.
pI,  mol wt., hydrophobicity, etc).  I'm afraid the only answer is to
try it find out.  Good luck.  -- D.G. Spinella



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