Complex-specific antibodies and antibody fragments and its use
The present invention provides antibodies and fragments thereof that specifically detect the complex of a specific cognate antigen-binding moiety, in particular antibodies, and its antigen. The antibodies of the present disclosure do not bind either said cognate antigen binding moiety or said antigen alone and this can be used e.g. to directly detect antigen-bound antigen-binding moieties. Further disclosed are methods for production and use of said antibodies and antibody fragments.
1 . A method of detecting, quantifying or monitoring a level of a complex of a specific cognate antibody or fragment thereof and its antigen, in a sample, the method comprising the steps of:
(a) screening a library of antibodies or antibody fragments against the complex of the specific cognate antibody and its antigen in the presence of the unbound antigen and an antibody different from the specific cognate antibody that has the same isotype as the specific cognate antibody,
(b) isolating a complex of the specific cognate antibody and its antigen and the bound antibodies or antibody fragments,
(c) identifying and isolating an isolated monoclonal antibody, or functional fragment thereof which specifically binds to a complex of a specific cognate antibody and its antigen and does not bind either said cognate antibody or said antigen alone;
(d) contacting said sample with the identified, isolated monoclonal antibody or fragment thereof,
(e) detecting said isolated monoclonal antibody or fragment bound to said complex, and
(f) correlating said isolated monoclonal antibody or fragment thereof bound to said complex with the concentration of the antigen-bound cognate antibody,
wherein said sample is obtained from an experimental animal or a human and said complex is detected using EIA, ELISA, RIA, indirect competitive immunoassay, direct competitive immunoassay, non-competitive immunoassay, sandwich immunoassay, agglutination assay or mesoscale discover (MSD).
2 . The method of claim 1 wherein said isolated monoclonal antibody or fragment thereof binds to at least parts of the CDR regions of the cognate antibody.
3 . The method of claim 1 wherein said isolated monoclonal antibody or fragment thereof is a human or humanized antibody or fragment thereof.
4 . The method of claim 1 wherein said cognate antibody or fragment thereof is a therapeutic antibody or a therapeutic antibody fragment.
5 . The method of claim 1 wherein said cognate antibody or fragment thereof is a diagnostic antibody or a diagnostic antibody fragment.
6 . The method of claim 1 wherein the antigen is a cytokine or a receptor.