AI Article Synopsis

  • Researchers developed cattle-derived ultralong antibodies targeting the epidermal growth factor receptor (EGFR) using a combination of cattle immunization and yeast display techniques.
  • These antibodies were able to effectively induce cytotoxicity in EGFR-overexpressing tumor cells and inhibit EGFR-mediated signaling without competing with existing drugs like Cetuximab or Matuzumab.
  • A unique aspect of the study highlighted that certain parts of the antibodies, called CDR-H3 knobs, could act independently to bind EGFR, indicating potential for future biotechnological applications after optimization.

Article Abstract

In this work, we have generated epidermal growth factor receptor (EGFR)-specific cattle-derived ultralong CDR-H3 antibodies by combining cattle immunization with yeast surface display. After immunization, ultralong CDR-H3 regions were specifically amplified and grafted onto an IGHV1-7 scaffold by homologous recombination to facilitate Fab display. Antigen-specific clones were readily obtained by fluorescence-activated cell sorting (FACS) and reformatted as chimeric antibodies. Binning experiments revealed epitope targeting of domains I, II, and IV of EGFR with none of the generated binders competing with Cetuximab, Matuzumab, or EGF for binding to EGFR. Cattle-derived chimeric antibodies were potent in inducing antibody-dependent cell-mediated cytotoxicity (ADCC) against EGFR-overexpressing tumor cells with potencies (EC killing) in the picomolar range. Moreover, most of the antibodies were able to significantly inhibit EGFR-mediated downstream signaling. Furthermore, we demonstrate that a minor fraction of CDR-H3 knobs derived from generated antibodies was capable of independently functioning as a paratope facilitating EGFR binding when grafted onto the Fc part of human IgG1. Besides slightly to moderately diminished capacities, these engineered Knobbodies largely retained main properties of their parental antibodies such as cellular binding and triggering of ADCC. Hence, Knobbodies might emerge as promising tools for biotechnological applications upon further optimization.

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Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8573386PMC
http://dx.doi.org/10.3389/fimmu.2021.742418DOI Listing

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