The chemical synthesis of knob domain antibody fragments
| Autor: | Yalan Tang, Jean M. H. van den Elsen, Phil Stanley, Maisem Laabei, Toska Wonfor, Callum Joyce, Richard D. Taylor, Kenneth Saunders, Robert J. Broadbridge, Alastair D. G. Lawson, Sebastian Kelm, Gregory Bogle, Matthew Duncan Selby, Douangsone Vadysirisack, Alex Macpherson, Kevin Brady, Jiye Shi, Richard J. Franklin, James R. Birtley, John Horton, Adam Hold, Monika-Sarah E. D. Schulze |
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| Rok vydání: | 2021 |
| Předmět: |
Male
Models Molecular Protein Folding viruses Peptide Complementarity determining region Computational biology Peptides Cyclic Biochemistry Rats Sprague-Dawley Protein Domains Palmitoylation Tandem Mass Spectrometry parasitic diseases Animals Amino Acid Sequence Immunoglobulin Fragments Solid-Phase Synthesis Techniques chemistry.chemical_classification Drug discovery Rational design General Medicine Biological product Complementarity Determining Regions Amino acid chemistry Thermodynamics Molecular Medicine Cattle Paratope Protein Binding |
| Popis: | Cysteine-rich knob domains found in the ultralong complementarity determining regions of a subset of bovine antibodies, are capable of functioning autonomously as 3-6 kDa peptides. While they can be expressed recombinantly in cellular systems, in this paper we show that knob domains are also readily amenable to chemical synthesis, with a co-crystal structure of a chemically synthesised knob domain in complex with antigen showing structural equivalence to the biological product. For drug discovery, following immunisation of cattle, knob domain peptides can be synthesised directly from antibody sequence data, combining the power and diversity of the bovine immune repertoire with the ability to rapidly incorporate non-biological modifications. We demonstrate that, through rational design with non-natural amino acids, paratope diversity can be massively expanded, in this case improving the efficacy of an allosteric peptide. As a potential route to further improve stability, we also performed head-to-tail cyclisation, exploiting the unusual proximity of the N- and C-termini to synthesise functional, fully cyclic antibody fragments. Lastly, we highlight the stability of knob domains in plasma and, through pharmacokinetic studies, use palmitoylation as a route to extend the plasma half-life of knob domains in vivo. This study presents an antibody-derived medicinal chemistry platform, with protocols for solid-phase synthesis of knob domains; together with characterisation of their molecular structures, in vitro pharmacology and pharmacokinetics. |
| Databáze: | OpenAIRE |
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