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Anti-Small Molecule Immune Complex Antibody Development
There are two basic forms of immunoassay, competitive and non-competitive, and the main application for small molecule detection is the competitive form of immunoassay. Non-competitive immunoassay theoretically has higher sensitivity and linear range than the competitive type, and the content of analyte and the detection signal are negatively correlated in competitive immunoassay, while positively correlated in non-competitive immunoassay. Figure 1 visualizes the superiority of non-competitive immunoassay in terms of detection sensitivity.
Fig.1 Comparison of sensitivity between non-competitive and competitive analysis
Small molecules are difficult to achieve non-competitive immunoassay due to their small molecular weight (<1000 Da) and single epitope, how can non-competitive immunoassay be achieved? Researchers have discovered a class of antibodies called anti-metetype antibodies, which can recognize new epitopes formed by the binding of small molecules to antibodies.
When a small molecule binds to an antibody, more than 85% of the antigen is encapsulated close to the surface area, and only a very small portion forms a heterotypic epitope with the antibody, making the preparation of heterotypic antibodies particularly difficult. Small molecule antibodies are generally derived from murine monoclonal antibodies, and if the mouse is then immunized with a murine anticomplex, there is essentially no immune response. Routinely, it is necessary to immunize rabbits with murine anticomplexes, but the development of rabbit monoclonal antibodies, whether it is hybridoma, phage display or SINGLE B, faces the problem of low efficiency.
Based on the HTS-Smart Select platform, NBbiolab can generate anti-idiotypic antibodies by immunizing small-molecule complex antibodies, then construct yeast display libraries, and directly sort anti-idiotypic antibodies through flow analysis. The whole process can be finely controlled to efficiently screen high-affinity and high-specific antibodies.
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