ELISA with virus as antigen - (Nov/06/2006 )

HI there

I am trying to check my protein level expressed in plants by sandwitch elisa. In short, I coat the polyclonal antibody at 4 degree, overnight then wash and blocking with 1%BSA in PBS, wash and then coat the plant extract and the virus for positive control( Virus, the surface protein of which has been expressed in plants ) ( 2000ng/ml to 500ng/ml). and then primary antibody and conjugated antibody.

while standarizing with only positive control ( taking commercial virus ) i couldnt find any linearity range.

can whole virus be used in sandwitch elisa ? can we estimate the virus surface protein concentration without disrupting the virus?( using Bradford Dye , biorad).

thanks

Have you tried varying your coat concentration or using a primary-enzyme conjugate instead of two steps? Both have improved linearity for me in the past.