Abstract
BackgroundIn certain cases, anti-idiotypic antibodies that recognize an antigen-combining site of an antibody can mimic the structure and/or function of certain nominal antigens. This feature makes them particularly useful if conventional experimental approaches fail to fulfil expectations, especially when the molecule of interest is infectious, toxic or difficult to isolate and purify. We suggest the application of an anti-idiotype concept to the field of prion biology, with the aim of evoking a humoral immune response against the pathological isoform of the prion protein (PrPSc). Different ways to induce anti-idiotypic responses were studied in mice and chickens using various forms of V5B2, a PrPSc-specific monoclonal antibody we have described previously.ResultsThe preparation of anti-idiotypic monoclonal antibodies was achieved with well-defined strategies of immunization, selection and subsequent characterization. Our results demonstrate that it is possible to induce a strong anti-idiotypic immune response against the V5B2 monoclonal antibody in both xenogeneic and syngeneic experimental systems. From the competition seen between polyclonal and monoclonal anti-idiotypic antibodies and the original immunogen, the P1 peptide, and even more importantly, the ultimate target antigen, PrPSc, we conclude that selected antibodies bind to the antigen-combining site of the V5B2 monoclonal antibody and might even resemble the PrPSc-specific epitope. The involvement of both antigen-combining sites in the interaction between V5B2 and the most promising monoclonal anti-idiotypic antibody was further supported by molecular docking.ConclusionThe results of the present study not only provide an example of the successful production of Ab2 monoclonal antibodies based on a well planned strategy for selection, but should also provide a new experimental approach that is applicable to the field of prion diseases.
Highlights
In certain cases, anti-idiotypic antibodies that recognize an antigen-combining site of an antibody can mimic the structure and/or function of certain nominal antigens
Each line represents the mean absorbance of 5 individual chicken sera ± standard deviation. (B) Competition assay for polyclonal chicken sera: for the competition assay, V5B2 was pre-incubated with the chicken sera that had been previously immunized with V5B2, and the subsequent binding ability of V5B2 to the P1 peptide was measured by ELISA
Each column represents the mean inhibition of V5B2 binding to the P1 peptide from the 4 individual chicken sera with the highest antibody titres ± standard deviation. (C) Competition assay for immunity purified chicken IgY (Ch5): each column represents the mean inhibition of V5B2 binding to the P1 peptide of at least three experiments ± standard deviation
Summary
Anti-idiotypic antibodies that recognize an antigen-combining site of an antibody can mimic the structure and/or function of certain nominal antigens. This feature makes them useful if conventional experimental approaches fail to fulfil expectations, especially when the molecule of interest is infectious, toxic or difficult to isolate and purify. Each antibody constitutes a small set of idiotopes that form its own idiotype. Private idiotopes have been shown to be associated with the complementarity-determining regions (CDRs), which, in addition to various rearrangements of V-(D)-J gene segments, reflect random somatic mutations and/or N-region additions with a low probability of repetition in another individual. A single idiotope can stretch over a part of the CDR and a part of the framework region, as well as over both the light and heavy chain residues
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