Monoclonal antibodies specific for the native, disease-associated isoform of the prion protein.
Reviewing the circumstances that have led to the first monoclonal antibody against the disease-associated form of PrP, we consider the availability of PrP knockout mice and recombinant PrP, as well as a reliable conformational screening protocol as being important prerequisites for a successful immunization approach. When considering presenting an antigen to a mouse with the goal of obtaining specific monoclonal antibodies against a misfolded or aggregated form of a host protein, it is desirable to increase the definition of a subtle conformational difference. This can be achieved by immunizing an antigen knockout mouse that has not developed self-tolerance against the respective antigen. Furthermore, if conformational isoforms and/or oligomeric forms of a protein sequence are understood to exist in an equilibrium, high and pure amounts of recombinant protein may increase the likelihood that a particular population of protein conformation passes an antigenic threshold necessary to start an immunogenic response. Pulling out the monoclonal antibodies by correct screening is essential. Screening against the pure misfolded or aggregated protein is often complicated by its poor solubility and hence the ability to immobilize. In the present case, immobilization of disease-associated PrP on nitrocellulose had been established as a conformation-sensitive screening method, allowing to "freeze" PrP in its distinguishable, disease-associated conformation. We are cautious to generalize conclusions of how to assess the generation of monoclonal antibodies against these particular protein isoforms to other diseases of protein misfolding and/or aggregation, but ultimately the present approach may inspire respective experiments.
Korth
C
C
Department of Neurology, University of California, San Francisco 94143-0518, USA.
Streit
P
P
Oesch
B
B
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005260:Female; D006825:Hybridomas; D015151:Immunoblotting / Q000379:methods; D051379:Mice; D018345:Mice, Knockout; D008969:Molecular Sequence Data; D019151:Peptide Library; D011233:Precipitin Tests; D011328:Prions / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D020033:Protein Isoforms / Q000276:immunology / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000276:immunology / Q000302:isolation & purification
D000911:Antibodies, Monoclonal; D019151:Peptide Library; D011328:Prions; D020033:Protein Isoforms; D011994:Recombinant Proteins