Figure EV5. TR‐FRET for the detection of wtPrP^{S c} in prion‐infected CAD5 Prnp ^+/+ after treatment with a panel of Fabs.

• A
  TR‐FRET for the detection of PrP^Sc in prion‐infected CAD5 Prnp ^+/+ cells after treatment with selected anti‐PrP Fabs. Treatment of the cells with Fab71 (OR_51‐91), but not with Fab3 (CC1_23‐50) or Fab29 (GD), significantly reduced PrP^Sc levels compared to untreated, prion‐infected cells.
• B
  Treatment of prion‐infected CAD5 Prnp ^+/+ cells with Fab100 (OR_51‐91 binder), but not with Fab83 (CC1_23‐50 binder), significantly reduced wtPrP^Sc levels compared to untreated prion‐infected CAD5 Prnp ^+/+ cells. The dilution range (3 × 10⁻⁵% to 1% w/v) of prion‐infected BH is shown on the right site to indicate the linear range of the assay. Negative controls: NBH and CAD5 Prnp ^−/− cells. Here and in (A), the FRET signal of untreated CAD5 Prnp ^−/− cells was set to zero. The FRET anti‐PrP antibody pair POM19‐Eu and POM1‐APC was used for detection.

Data information: For prion‐infected CAD5 Prnp ^+/+ cells, the assay was performed in biological triplicates represented as four technical replicates for Fab71, Fab3, Fab100, and Fab83. For CAD5 Prnp ^+/+ cells that were either untreated, exposed to NBH, or treated with Fab29, the assay was performed in biological duplicates represented as four technical replicates. For CAD5 Prnp ^−/− cells, one biological replicate was assayed as four technical replicates. n = 4–12. Each dilution of prion‐infected BH is represented in technical duplicates. Data presented in dot plots represent the mean ± sem. One‐way ANOVA and Bonferroni's multiple comparisons were used for statistical analysis; *P < 0.05; **P < 0.01.