Production of Designer VHH-Based Antibodies in Plants

Methods Mol Biol. 2022:2446:205-230. doi: 10.1007/978-1-0716-2075-5_10.

Abstract

Simplified monoclonal antibodies can be produced by fusing a VHH or nanobody, derived from camelid heavy-chain-only antibodies to the Fc domain of either IgG (VHH-IgG), IgA (VHH-IgA), or IgY (VHH-IgY). These recombinant antibodies are encoded by a single gene and their production can be easily scaled up in plants. This chapter contains methods for Gateway cloning of VHH-Fc fusions into the binary T-DNA vectors pEAQ-HT-DEST1 and pPhasGW, electroporation of Agrobacterium with the resulting constructs, transient antibody expression in Nicotiana benthamiana leaves, and stable antibody expression in Arabidopsis thaliana seeds. The properties of chimeric VHH-based antibodies produced in plants enable novel passive immunization treatments, such as in-feed oral delivery or intravenous injection.

Keywords: Agrobacterium-mediated transformation; Nanobody; Recombinant antibody production; Transgenic plants; Transient expression; VHH.

MeSH terms

  • Antibodies, Monoclonal / genetics
  • Antibodies, Monoclonal / metabolism
  • Immunoglobulin Heavy Chains / genetics
  • Immunoglobulin Heavy Chains / metabolism
  • Nicotiana / genetics
  • Nicotiana / metabolism
  • Plants, Genetically Modified / genetics
  • Plants, Genetically Modified / metabolism
  • Single-Domain Antibodies* / genetics
  • Single-Domain Antibodies* / metabolism

Substances

  • Antibodies, Monoclonal
  • Immunoglobulin Heavy Chains
  • Single-Domain Antibodies