Creative Diagnostics employs three approaches to generate monoclonal anti-idiotype antibodies (anti-IDs).
The first approach is to raise hybridoma clones in rats or mice. Any species of antibodies, whole IgG, scFv, Fab or F(ab’)2, that preserve the antibody activity can be used for anti-IDs development. To enrich the antibodies that target the antigen-binding site, we usually cut the target antibody into Fab fragments, which could be used as immunogen (Antibody Fragmentation service). After screening against the immunogen, we can use isotype matching control antibodies and related immunoglobulin/proteins to do counter-selection. In this case, only the anti-IDs with the correct specificity are expanded. We further run corresponding assays to make sure the anti-IDs’ lack of cross reactivity to non-specific antibodies and receptors. Additionally, our team can develop ELISA assays to support your studies, and set up neutralization and blocking assay to identify functional ligand blocking antibodies/receptors.
The second approach is to isolate V-regions specific antibodies from our human/mouse antibody libraries. For anti-IDs generation, screening is carried in the presence of isotype-matching control antibodies as blockers, to avoid enrichment of antibodies that bind to other regions of the target antibody. Additionally, selection is carried out in the presence of serum of the corresponding species and related immunoglobulin /proteins, which can avoid matrix effects in the final assay and ensure idiotope specificity. We offer these this service option at Creative BioLabs.
The third approach is using target antibody in scFv form or using a peptide designed according to the sequences of the variable domains as the immunogen. We only need to know the amino acid sequence of the variable regions of the target antibody to do so.
Creative Diagnostics has successfully developed a panel of high affinity and specificity monoclonal anti-IDs for our customers. We could develop the antibodies that work in your end application, concerning the items from routine screening, Elisa assay, immunohistochemistry, western blot, flow cytometry to custom functional assays. For more information, please check our Antibody Labeling and contact assay development service.