Anti-Prion protein [Nb484], Human IgG1-Fc Fusion,

Absolute Antibody

Anti-Prion protein [Nb484], Human IgG1-Fc Fusion,

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This antibody is specific for major prion proteins, with a demonstrated specificity for a discontinuous epitope on the human prion protein (HuPrP). It binds to specific structural elements and loops within HuPrP, particularly the beta0-beta1 loop (residue

The original version of this antibody (alpaca VHH) bound mouse and human prion proteins (MoPrP and HuPrP, respectively) as demonstrated by SPR binding assays with a Kd of 40 nM, 50 nM, 9.54 nM, and 7 microM for MoPrP(23-230), MoPrP(89-230), HuPrP(90-231), and HuPrP(23-144), respectively. The addition of this antibody to MoPrP(23-230) extended the lag phase of fibrillization by about 40 hours in an amyloid seeding assay (ASA), indicating that the interaction of this antibody with the full-length MoPrP inhibited the formation of prion protein infectious scrapie agent (PrPSc)-like aggregates. This antibodys ability to inhibit prion propagation was further confirmed by treating scrapie-infected murine cells (ScGT1) with different antibody concentrations. The treatment resulted in a dramatic, dose-dependent reduction in PrPSc levels, as measured by PK assay and Western blotting. At a concentration of 3.5 microM, PrPSc levels were no longer detectable, and this effect persisted even after removing the antibody from the medium. This antibody was complexed with HuPrP(23-231) and HuPrP(90-231), and their crystal structure was examined. The binding of this antibody stabilizes the conformation of the human prion protein, particularly the beta2-alpha2 loop, which is known to be flexible in the native structure but becomes rigid upon binding (Abskharon et al., 2014; PMID: 24400836). The binding of this antibody to recombinant PrP (recPrP) and recPrPSc was evaluated using ELISA. It was found to have a high affinity for recPrP, as previously reported, but its binding to the infectious recPrPSc is minimal, indicating that the antibody exerts its inhibitory effect on prion propagation through binding to the non-infectious recPrP. The binding of this antibody to recPrP was further supported by the results of the discontinuous iodixanol density gradient floatation assay. This antibody was crystallized with MoPrP(89-230) at pH 6.0 and 8.0 and alone (free form). This antibody inhibited prion conversion by competitively binding to the hydrophobic region of MoPrP, thereby blocking the interaction with cofactors essential for prion propagation, as evidenced by Protein Misfolding Cyclic Amplification (PMCA) assays and lipid binding studies (Abskharon et al., 2019; PMID: 31815959). This antibody recognizes and binds the beta2-alpha2 loop epitope across various HuPrP structures; the binding mechanisms of this antibody to different variants of HuPrP, including wild-type, E219K polymorphism, and V210I mutation, were studied using molecular dynamics simulations and experimental data. It was found to recognize flexible epitopes within the beta2-alpha2 loop of HuPrP through a conformational selection model, stabilizing diverse conformations present in these variants. Despite structural differences among HuPrP variants, the antibody exhibited high-affinity binding, emphasizing its ability to interact with disordered regions crucial for amyloidogenic protein recognition (Mollica & Giachin, 2023; PMID: 36580661).

Functional Assay, Western Blot, ELISA, Other Application

Research Use Only

Monoclonal

Nb484

PRNP

prion protein

alternative prion protein; AltPrP; ASCR; CD230; CD230 antigen; CJD; GSS; KURU; major prion protein; p27-30; prion-related protein; PRIP; PrP; PrP27-30; PrP33-35C; PrPc

Human

IgG1

Major prion protein

This is a chimeric antibody, developed from the original camelid VHH antibody, specifically engineered for enhanced compatibility with existing reagents, assays, and techniques.

Human, Mouse

-20°C,2°C to 8°C

12352203