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

Abstract

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.

Graphical Abstract

Scientific illustration outlining steps involved in the workflow of a proteomics-only sequencing platform for antibodies from goats

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.

Key Takeaways

  • pAb have been highly sought after due to their biophysical properties and high affinity to a target
  • It has been difficult to completely capture pAb properties without compromising sensitivity and/or reproducibility
  • Protein sequencing of pAbs, when combined with recombinant expression, can yield a collection of rAbs that perform as well or better than the original pAb
  • Full sequence coverage and correct chain pairing of mAbs directly from a protein sample is now possible

Talk to Our Scientists.

We Have Sequenced 6000+ Antibodies and We Are Eager to Help You.

Through next generation protein sequencing, Rapid Novor enables reliable discovery and development of novel reagents, diagnostics, and therapeutics. Thanks to our Next Generation Protein Sequencing and antibody discovery services, researchers have furthered thousands of projects, patented antibody therapeutics, and developed the first recombinant polyclonal antibody diagnostics.

Talk to Our Scientists.

We Have Sequenced 6000+ Antibodies and We Are Eager to Help You.

Through next generation protein sequencing, Rapid Novor enables timely and reliable discovery and development of novel reagents, diagnostics, and therapeutics. Thanks to our Next Generation Protein Sequencing and antibody discovery services, researchers have furthered thousands of projects, patented antibody therapeutics, and ran the first recombinant polyclonal antibody diagnostics

Talk to our scientists. We have sequenced over 6000 antibodies and we are eager to help you.