D. Perez-Witzke1, T. Nunez de Villavicencio Diaz2, N. Moore1, T. Le Bihan2, N. Hutchings1, R. Sapkota3 , M. Mobley4 , M. Xie2, B. Ma2, and M. Fiebig1
1 Absolute Antibody, Redcar, UK
2 Rapid Novor, Inc, Kitchener, Canada
3 Shikar Biotech, Kathmandu, Nepal
4 Everest Biotech Ltd, Bicester, UK
Circulating in blood is a multitude of biologically important antibodies. These pools of polyclonal antibodies (pAb) are invaluable sources for drug discovery against various diseases, and for the development of robust immunoreagents for diagnostics, and research. However, accessing the richness of clinically relevant pAb remains difficult due to batch-to-batch variability, and shortcomings in available cell-based discovery. Herein, we present the first report of the de novo sequencing of monoclonal antibodies (mAbs) from a goat pAb without cell or nucleic material input. The decoded mAb sequences were recombinantly expressed and analyzed with enzyme-linked immunosorbent assays (ELISA). The discovered antibodies faithfully recapitulated the specific binding affinity of the polyclonal antibody.
Steps involved in this case study, where the REpAb® antibody discovery platform (proteomics-only option) was used for sequencing a caprine-harvested and purified anti-AIF1 polyclonal antibody mixture, followed by recombinant expression and binding affinity testing via ELISA.