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Biochem Biophys Res Commun. 2018 Feb 5;496(2):614-620. doi: 10.1016/j.bbrc.2018.01.051. Epub 2018 Jan 9.

Production and characterization of a novel site-specific-modifiable anti-OX40-receptor single-chain variable fragment for targeted drug delivery.

Author information

1
Department of Computational Biology and Medical Sciences, Graduate School of Frontier Sciences, The University of Tokyo, Tokyo, Japan.
2
Department of Bioengineering, School of Engineering, The University of Tokyo, Tokyo 113-8656, Japan.
3
Institute of Medical Science, The University of Tokyo, Tokyo 108-8639, Japan.
4
Department of Immunology, Graduate School of Medicine, University of the Ryukyus, Okinawa, 903-0215, Japan.
5
Department of Bioengineering, School of Engineering, The University of Tokyo, Tokyo 113-8656, Japan; Institute of Medical Science, The University of Tokyo, Tokyo 108-8639, Japan. Electronic address: tsumoto@ims.u-tokyo.ac.jp.
6
Department of Computational Biology and Medical Sciences, Graduate School of Frontier Sciences, The University of Tokyo, Tokyo, Japan; Hematology/Oncology, Research Hospital, Institute of Medical Science, The University of Tokyo, Tokyo, Japan.
7
Department of Computational Biology and Medical Sciences, Graduate School of Frontier Sciences, The University of Tokyo, Tokyo, Japan; Department of Advanced Medical Innovation, St. Marianna University School of Medicine, Kanagawa, Japan. Electronic address: tnabe@ims.u-tokyo.ac.jp.

Abstract

OX40 receptor (tumor necrosis factor receptor superfamily, member 4; CD134) is a T-cell co-stimulatory molecule that plays an important role in T-cell activation and survival. OX40 receptor is activated by its ligand, OX40L; and modulation of the OX40-OX40L interaction is a promising target for the treatment of autoimmune diseases and cancers. Here, we generated a high-affinity anti-OX40 single-chain variable fragment carrying a C-terminal cysteine residue (scFvC). Physicochemical and functional analyses revealed that the scFvC bound to OX40-expressing cells and was internalized via OX40-mediated endocytosis without inducing phosphorylation of IκBα (nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor, alpha), an important complex in the classical NFκB (nuclear factor kappa-light-chain-enhancer of activated B cells) signaling pathway. In addition, mutation of the 36th cysteine residue in variable region of light chain enabled site-specific chemical modification to carboxy terminal cysteine and improved the thermal stability of the scFvC. These results suggest that this novel high-affinity anti-OX40 scFvC may be useful as a transporter for targeted delivery of small compounds, proteins, peptides, liposomes, and nanoparticles, into OX40-expressing cells for the treatment of autoimmune diseases and cancers.

KEYWORDS:

Antibody drug conjugate; Drug delivery; Internalization; OX40; Single chain antibody(scFv); Thermal stability of antibody

PMID:
29330050
DOI:
10.1016/j.bbrc.2018.01.051
[Indexed for MEDLINE]

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