AI Article Synopsis
- Antibody-based cancer therapies, particularly bispecific antibody-drug conjugates, are advancing quickly in the pharmaceutical industry, enhancing immunotherapy's effectiveness.
- Miniaturized antibody fragments like diabodies, nanobodies, and scFvs show great potential for targeting and penetrating tumor tissues in cancer treatments.
- This study developed a versatile scFv OKT3 antibody using E. coli, incorporating a unique amino acid for efficient 'click chemistry' conjugation, intending for applications in controlled anti-T cell therapies and cancer imaging.
Article Abstract
Antibody-based cancer therapies have been evolving at a rapid pace in the pharmaceutical market. Bispecific antibody-drug conjugates that engage immune cells to target and kill cancer cells with precision have inspired the development of immunotherapy. Miniaturized antibody fragments such as diabodies, nanobodies, or single-chain variable fragments (scFvs) hold great promise as antibody-drug conjugates as they specifically target tumor tissue and can penetrate it. Here, we optimized the soluble periplasmic expression of the scFv OKT3 comprising the variable V and V domains of the mouse anti-human CD3 antibody muromonab-CD3 (trade name Orthoclone OKT3) in E. coli. By an expansion of the genetic code, we site-specifically incorporated the reactive non-canonical amino acid N-((2-azidoethoxy)carbonyl)-L-lysine (AzK) into scFv OKT3 using an orthogonal pyrrolysyl-tRNA synthetase/tRNA pair. To confirm the AzK incorporation and to demonstrate the accessibility of the reactive azide group, we conjugated a fluorophore to scFv OKT3 AzK variants by copper-free strain-promoted alkyne-azide cycloaddition ('click chemistry'). The scFv OKT3 wild type and the AzK variants bound T cells at nanomolar concentrations. In this study, a 'ready-to-click' scFv OKT3 was successfully developed for future applications, e.g. as controlled anti-T cell antibody-drug conjugate or bispecific T cell engager and for imaging immune T cell migration in cancers.
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| http://dx.doi.org/10.1016/j.nbt.2023.05.007 | DOI Listing |
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Article Synopsis
- Antibody-based cancer therapies, particularly bispecific antibody-drug conjugates, are advancing quickly in the pharmaceutical industry, enhancing immunotherapy's effectiveness.
- Miniaturized antibody fragments like diabodies, nanobodies, and scFvs show great potential for targeting and penetrating tumor tissues in cancer treatments.
- This study developed a versatile scFv OKT3 antibody using E. coli, incorporating a unique amino acid for efficient 'click chemistry' conjugation, intending for applications in controlled anti-T cell therapies and cancer imaging.
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Department of Medicine, Division of Hematology and Oncology, University of Virginia Cancer Center, Charlottesville, VA, USA.
Bispecific antibody (BiAb)-armed activated T cells (BATs) comprise an adoptive T cell therapy platform for treating cancer. Arming activated T cells (ATC) with anti-CD3 x anti-tumour associated antigen (TAA) BiAbs converts ATC into non-major histocompatibility complex (MHC)-restricted anti-tumour cytotoxic T lymphocytes (CTLs). Binding of target antigens via the BiAb bridge enables specific anti-tumour cytotoxicity, Th1 cytokines release, and T cell proliferation.
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