Figure 4
From: Disease-Associated Prion Protein Elicits Immunoglobulin M Responses In Vivo
Polyclonal antibodies from mice immunized with PrPSc-Dynabeads recognize mouse rβPrP in ELISA. Anti-PrP responses were assayed in mice (n = 5 for each arm) immunized with RML (PrPSc) or Prn-Po/o brain homogenates adsorbed or not to antibody (ICSM or an isotype control mAb BRIC 126)-coated sheep anti-mouse Dynabeads-M450. PrP was adsorbed from 100 µL of a 1% PK-digested homogenate, or for arms without Dynabeads, 100 µL of a 1% homogenate emulsified in Freunds was used for immunization. Sheep anti-mouse Dynabeads-M450 coated with anti-PrP (ICSM 35) or an isotype-match control mAb (BRIC 126) were used to adsorb after proteinase K digestion. After omission of the Dynabeads, anti-PrP (ICSM 35) or isotype-match control mAb (BRIC 126) were used to bind RML (PrPSc) or Prn-Po/o brain homogenates. Serum were assayed at 1:100 dilution. The assay negative control (Prn-Po/o mouse serum) gave optical density (OD) values of 0.1 ± 0.03.