Due to their ability to bind biomolecules specifically and tightly, antibodies are highly sought after as invaluable reagents for basic biological research, clinical diagnostics, and therapeutic treatments. Consequently, antibody discovery campaigns have become increasingly appealing to identify, characterize, and optimize antibodies with novel targets and functionalities. In a typical discovery workflow, antibody candidates are generated by confirming the interaction between the antibodies and the target of interest, leading to potential ‘hits’. At this stage, it is commonplace to have a large number of candidates, but how does one move forward to select lead antibody molecules for further research and development?
Here, we demonstrate the application of kinetic analysis by surface plasmon resonance (SPR) in a proof-of-concept antibody discovery workflow. Using our REpAb® Antibody Discovery Platform, we sequenced the population of polyclonal antibodies directly from an alpaca’s serum. From this population, six antibodies with strong polyclonal binding activity were screened, validated, and analyzed against two targets through kinetic profiling by SPR. As a result, four lead candidates were identified based on their kinetic profiles indicating their highly unique specificities, selectivities, affinities, and other kinetic parameters (ie. on-rate and off-rate).
Advancements in de novo protein sequencing and proteomics techniques, like those applied here, make it possible to accelerate the discovery workflow by minimizing the need to produce the entire collection of potential candidate molecules. One of the many advantages to this approach is the capability to prioritize candidates with valuable attributes in manageable volumes. Strategies like these have the potential to speed up the discovery process, making these critically important reagents more accessible.