Mouse Anti-PrP Recombinant Antibody (clone sc4.1) (CAT#: VS-0623-WK5)

Figure 1 Western blot analysis of anti-PrP scFv protein (clone 4.1A, lane 2).

Supernatants derived from cells transfected with PSA (lane 3) or GFP (lane 1) served as positive and negative controls.

Figure 2 Dot blot analysis of anti-PrP scFv protein (clone 4.1A).

Recombinant PrP (PrPREC; 200 ng) was spotted onto nitrocellulose and incubated with medium derived from RD-4 cells stably expressing anti-PrP scFv (clone 4.5 and clone 4.1), GFP, and PSA.

Figure 3 Affinity-purified sc4.1 resulted in concentration-dependent binding to PrPᒼ (an abnormal form of the cellular prion protein) as assessed by ELISA.

Figure 4 Inhibition analysis of scFv (clone 4.1) binding to Recombinant PrP by ELISA.

Pretreatment of Recombinant PrP-coated ELISA plates with increasing amounts of monoclonal antibody 6H4 (recognizing mouse PrP144-152) inhibited binding of 4 μg purified scFvs (clone 4.1) to PrPREC (squares), while anti-PrP antibody POM2 (which recognizes an amino-terminal epitope of PrPC) did not alter PrP reactivity of sc 4.1 (triangles).

Figure 5 Inhibition analysis of scFv (clone 4.1) binding to Recombinant PrP by WB.

Monoclonal antibody 6H4, but not the IgG isotype control, antagonized binding to Recombinant PrP when administered prior to incubation with sc4.1.

Figure 6 Anti-PrP scFvs rescue chronically scrapie-infected ScN2a cells by In-cell Western assay.

Decrease in PrPSc content in ScN2a cells cocultured with stably transfected RD-4 cells producing large amounts of anti-PrP scFvs (4.1 high, in duplicate, lanes 1 and 2, upper row). Ethidium bromide staining demonstrates the presence of equal amounts of cells per well upon transfer onto nitrocellulose membrane (lower row).

Figure 7 PrP^Sc load after 12 days of treatment with anti-PrP-scFv.

After 3 weeks of coculturing ScN2a cells with anti-PrP-scFv-producing RD-4 cells (clone 4.1), ScN2a cells were left in culture for three further passages (corresponding to 12 days) without further anti-PrP scFvs treatment. A strong reduction of PrPSc was observable in ScN2a cells that had previously been exposed to RD-4 cells producing high levels of anti-PrP scFv (clone 4.1 high, in duplicate; lanes 3 and 4).