Immunodetection of PrPSc oligomers/multimers with PRIOC mAbs by Sandwich ELISA.

(A). Principle of a newly developed specific and sensitive ELISA designed for the detection of PrPSc oligomers/multimers. (B). RML-infected brain homogenates were used to assess specific binding of PRIOC mAbs to PrPSc oligomers following proteinase K (PK) digestion. 50 µl of 5 µg/ml of PRIOC or anti-monomer antibody (AB) was used to coat the ELISA plate in coating buffer. RML-infected brain homogenate was added to the wells followed by a biotinylated PRIOC mAb or anti-monomer antibody (AB). The sandwich format of the assay has established the specificity of PRIOC antibody for post-PK PrPSc oligomers. Error bars represent the mean antibody level derived from n = 4 wells.