Antibody pairs are two different antibodies generated against different epitopes or binding regions, but with the same target. This unique property allows the simultaneous binding to the same target against two different epitopes, which present great potential in biological research. Antibody pair can be used to detect and quantify interaction molecules in situ, ensure localization of sub-cellular events, reduce the risk of missing weak or transient interactions and improve detection specificity and sensitivity.
An antibody pair always includes a capture antibody and a detection antibody, and is applied in sandwich ELISA, IP-WB, and protein-protein interaction. Monoclonal antibodies can be spread used in all types of ELISAs and also work with a polyclonal antibody to improve the chance of capturing antigen from a complex solution. Polyclonal antibodies present all the available epitopes in any given antigen but must be tested and validated thoroughly.
Fig.1 Schematic Representation of the Cascade Signal Amplification Strategy for Protein Detection. (Cheng, 2010)
Surface plasmon resonance (SPR) is a technique that allows label-free, optical monitoring of important kinetic information, such as the association and dissociation rates of antibodies. Using SPR it is possible to screen crude antibodies. When developing sandwich format assays for large molecular weight proteins it is essential to select appropriate antibody pairs for the capture and detection of the target analyte. Sandwich pairs are antibodies that are capable of simultaneously binding an antigen. Using SPR to select antibody pairs for use in LFIA saves time and can be largely automated for screening large antibody panels for their pairs.
A matched antibody pair for ELISA consists of two antibodies, a capture and a detection antibody. In a matched ELISA pair, each antibody is specific for a different and non-overlapping region or epitope of the antigen.
Rolling circle amplification (RCA) is emerging as a useful process for on-chip signal amplification and it is attractive for multiplexed microarray immunoassays. Recently, RCA-amplified protein chips have been established as concerns about specificity and sensitivity are being overcome by careful quality control and amplification technologies. RCA-amplified protein chips can select antibody pairs in various substrates including serum, plasma, and supernatants with high sensitivity, broad dynamic range and good reproducibility. The diagnostic utility of RCA-amplified protein chips has been shown for multiplexed allergen testing.
References
For Research Use Only.
