Programmable half-life and anti-tumour effects of bispecific T-cell engager-albumin fusions with tuned FcRn affinity

Commun Biol. 2021 Mar 8;4(1):310. doi: 10.1038/s42003-021-01790-2.

Abstract

Fc-less bispecific T-cell engagers have reached the immuno-oncology market but necessitate continual infusion due to rapid clearance from the circulation. This work introduces a programmable serum half-life extension platform based on fusion of human albumin sequences engineered with either null (NB), wild type (WT) or high binding (HB) FcRn affinity combined with a bispecific T-cell engager. We demonstrate in a humanised FcRn/albumin double transgenic mouse model (AlbuMus) the ability to tune half-life based on the albumin sequence fused with a BiTE-like bispecific (anti-EGFR nanobody x anti-CD3 scFv) light T-cell engager (LiTE) construct [(t½ 0.6 h (Fc-less LiTE), t½ 19 hours (Albu-LiTE-NB), t½ 26 hours (Albu-LiTE-WT), t½ 37 hours (Albu-LiTE-HB)]. We show in vitro cognate target engagement, T-cell activation and discrimination in cellular cytotoxicity dependent on EGFR expression levels. Furthermore, greater growth inhibition of EGFR-positive BRAF mutated tumours was measured following a single dose of Albu-LiTE-HB construct compared to the Fc-less LiTE format and a full-length anti-EGFR monoclonal antibody in a new AlbuMus RAG1 knockout model introduced in this work. Programmable half-life extension facilitated by this albumin platform potentially offers long-lasting effects, better patient compliance and a method to tailor pharmacokinetics to maximise therapeutic efficacy and safety of immuno-oncology targeted biologics.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3T3 Cells
  • Animals
  • Antibodies, Bispecific / metabolism
  • Antibodies, Bispecific / pharmacokinetics*
  • Antineoplastic Agents, Immunological / metabolism
  • Antineoplastic Agents, Immunological / pharmacokinetics*
  • CHO Cells
  • Cricetulus
  • Drug Compounding
  • Female
  • HEK293 Cells
  • HT29 Cells
  • Half-Life
  • Histocompatibility Antigens Class I / metabolism*
  • Homeodomain Proteins / genetics
  • Humans
  • Jurkat Cells
  • Lymphocyte Activation / drug effects
  • MCF-7 Cells
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Neoplasms / drug therapy*
  • Neoplasms / immunology
  • Neoplasms / pathology
  • Proof of Concept Study
  • Protein Binding
  • Receptors, Fc / metabolism*
  • Recombinant Fusion Proteins / metabolism
  • Recombinant Fusion Proteins / pharmacokinetics
  • Serum Albumin, Human / genetics
  • Serum Albumin, Human / metabolism
  • Serum Albumin, Human / pharmacokinetics*
  • T-Lymphocytes / drug effects*
  • T-Lymphocytes / immunology
  • Tumor Burden / drug effects
  • Xenograft Model Antitumor Assays

Substances

  • Antibodies, Bispecific
  • Antineoplastic Agents, Immunological
  • Histocompatibility Antigens Class I
  • Homeodomain Proteins
  • Receptors, Fc
  • Recombinant Fusion Proteins
  • RAG-1 protein
  • Fc receptor, neonatal
  • Serum Albumin, Human