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Selection of antibodies using HuCAL® technology is performed in vitro. This enables greater flexibility for antibody generation than is possible using traditional methods based on the immunization of animals.
‘Guided Selection’ is the term we use for all selection strategies used for the identification of antibodies to specific epitopes or epitope areas. Two strategies are typically used for the generation of sandwich pair antibodies.
Fig. 1. Selection with a captured antigen. The antigen is captured by an antibody, previously isolated from the library. Panning is carried out on the complex to select for the detection antibody.
The standard panning project (strategy 1) generates Fab antibodies in as little as 8 weeks. A further 1-2 weeks is required to find the best combination of capture and detection antibodies. The second strategy requires an initial panning of the library to select the capture antibody (8 weeks, plus 1-2 weeks testing to find the best antibody) and a second panning to select the detection antibody (8 weeks), plus 1-2 weeks for testing to find the best antibody pair.
Depending on the availability of your target of interest, the project can also start with the sequence of your protein and include an antigen generation step, which can be done using gene synthesis and expression or peptide synthesis and coupling. An additional 5-10 weeks is needed for this depending on the route chosen.
Step 1: Antigen generation
Step 2: Antibody generation
Step 3: Sandwich ELISA testing
Fig. 2. Process flow for sandwich ELISA antibody pair selection using HuCAL technology.