biostudies-literatureMolgaard KNovo Nordisk FoundationConsejería de Educación, Juventud y Deporte, Comunidad de MadridNovo Nordisk Foundation (Denmark)Instituto de Salud Carlos IIIDanish Council for Independent Reserach - Medical SciencesEPFL FellowsMinisterio de Economía y CompetitividadNovo Nordisk Fonden1251-1260https://www.ebi.ac.uk/biostudies/studies/S-EPMC11028287biostudies-literatureUnknown67(8)The recruitment of T-cells by bispecific antibodies secreted from adoptively transferred, gene-modified autologous cells has shown satisfactory results in preclinical cancer models. Even so, the approach's translation into the clinic will require incremental improvements to its efficacy and reduction of its toxicity. Here, we characterized a tandem T-cell recruiting bispecific antibody intended to benefit gene-based immunotherapy approaches, which we call the light T-cell engager (LiTE), consisting of an EGFR-specific single-domain V_HH antibody fused to a CD3-specific scFv. We generated two LiTEs with the anti-EGFR V_HH and the anti-CD3 scFv arranged in both possible orders. Both constructs were well expressed in mammalian cells as highly homogenous monomers in solutioCancer immunology, immunotherapy : CIIBispecific light T-cell engagers for gene-based immunotherapy of epidermal growth factor receptor (EGFR)-positive malignancies.PMC11028287NNF14OC0011019665667S2010/BMD-2312CTQ2017-83810-RPI13/00090DFF-6110-00533Mikkelsen KAlvarez-Vallina LHarwood SLBonet JNunez-Prado NSanz LBlanco FJAlvarez-Cienfuegos AMolgaard KMerino NAlvarez-Mendez ACompte MfalseBispecific light T-cell engagers for gene-based immunotherapy of epidermal growth factor receptor (EGFR)-positive malignancies.The recruitment of T-cells by bispecific antibodies secreted from adoptively transferred, gene-modified autologous cells has shown satisfactory results in preclinical cancer models. Even so, the approach's translation into the clinic will require incremental improvements to its efficacy and reduction of its toxicity. Here, we characterized a tandem T-cell recruiting bispecific antibody intended to benefit gene-based immunotherapy approaches, which we call the light T-cell engager (LiTE), consisting of an EGFR-specific single-domain V_HH antibody fused to a CD3-specific scFv. We generated two LiTEs with the anti-EGFR V_HH and the anti-CD3 scFv arranged in both possible orders. Both constructs were well expressed in mammalian cells as highly homogenous monomers in solutio2018-01-01T00:00:00Z2018 Aug2026-06-02T01:43:56.859Z2025-04-06T03:15:38.702ZS-EPMC110282872986916810.1007/s00262-018-2181-5