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842 A novel agonistic anti-CD40 targeting strategy with an affinity peptide binding feature for antigen cargo functionality: improving peptide stability and T cell proliferation
  1. Ida Olsson,
  2. Mohamed Eltahir and
  3. Sara Mangsbo
  1. Uppsala University, Uppsala, Sweden


Background To induce a prominent anti-tumor T-cell response, a viral or tumor derived antigen epitope imbedded in a longer synthetic peptide (SLP) can be used, which also requires internalization and processing by antigen presenting cells (APCs) to enable T cell priming. Herein we present the design and evaluation of a CD40 targeting tetravalent bispecific antibody, binding peptides through affinity as an antibody-drug conjugate. APC activation as well as in vitro and in vivo T-cell proliferation studies demonstrate retained agonistic activity as well as improved T cell proliferation/expansion in vitro and in vivo, compared to non-linked peptide/antibody mixes.

Methods T-cell priming was evaluated with B3Z assay or a cytomegalovirus (CMV) model and displayed superior uptake to non-bound peptide in the co-stimulatory independent B3Z assay. In addition, intracellular peptide release in APCs was analysed using a unique quenching strategy displaying peptide release after around 4-6 hour post antigen.

Results Peptide stability in vitro, when bound to the antibody, was analysed by mass spectrometry and displayed prolonged peptide stability in serum, increasing the peptide half-life by 15 times in vitro (

Conclusions Data support that the novel delivery system can improve antigen targeting to dendritic cells, but can also provide a prolonged peptide half-life as well as a peptide delivery to APCs. Combined this improves the efficiency of both antigen delivery and CD40 agonistic functionality.

This is an open access article distributed in accordance with the Creative Commons Attribution 4.0 Unported (CC BY 4.0) license, which permits others to copy, redistribute, remix, transform and build upon this work for any purpose, provided the original work is properly cited, a link to the licence is given, and indication of whether changes were made. See:

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