Orientation of antigen binding sites in dimeric and trimeric single chain Fv antibody fragments
Abstract
Electron microscopy of dimeric and trimeric single chain antibody Fv fragments (scFvs) complexed with anti‐idiotype Fab fragments was used to reveal the orientation of antigen binding sites. This is the first structural analysis that discloses the multivalent binding orientation of scFv trimers (triabodies). Three different scFv molecules were used for the imaging analysis; NC10 scFv‐5 and scFv‐0, with five‐ and zero‐residue linkers respectively between the V H and V L domains, were complexed with 3‐2G12 anti‐idiotype Fab fragments and 11‐1G10 scFv‐0 was complexed with NC41 anti‐idiotype Fab fragments. The scFv‐5 molecules formed bivalent dimers (diabodies) and the zero‐linker scFv‐0 molecules formed trivalent trimers (triabodies). The images of the NC10 diabody‐Fab complex appear as boomerangs, not as a linear molecule, with a variable angle between the two Fab arms and the triabody‐Fab complexes appear as tripods.
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Diabodies can be expressed at high yields in bacteria and have been shown by crystallography to adopt several conformations.
“These molecules can be expressed at high yields in bacteria and have been shown by crystallography experiments to adopt several conformations. 35”
From Bispecific antibodies for cancer therapy: The light at the end of the tunnel? (2009) · cites this paper as [35]
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Bispecific antibodies for cancer therapy: The light at the end of the tunnel?
Chames, P., & Baty, D. (2009) · mAbs · doi:10.4161/mabs.1.6.10015 · Cited by 336 · see citing works
- Authors
- Lawrence, L. J., Kortt, A. A., Iliades, P., Tulloch, P. A., & Hudson, P. J.
- Affiliations
- Commonwealth Scientific and Industrial Research Organisation (AU)
- Corresponding author
- Hudson, Peter J.
- ORCID
- Hudson, Peter J.
- Venue
- FEBS Letters
- Publisher
- Wiley
- Published
- 1998-04-03
- Volume
- 425
- Issue
- 3
- Pages
- 479-484
- ISSN
- 0014-5793 (ISSN-L)
- 1873-3468
- OpenAlex
- W2013843528
- Cited by
- 50
- References
- 27
- Citations by year
- 2025: 1 · 2023: 1 · 2021: 1 · 2020: 2 · 2018: 1 · 2016: 1 · 2015: 1 · 2013: 2 · 2012: 4
- FWCI
- 6.85
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- FEBS Letters (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Health Sciences › Medicine › Radiology, Nuclear Medicine and Imaging › Monoclonal and Polyclonal Antibodies Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Biochemical and Structural Characterization
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Structure and Dynamics
- Keywords
- single-chain Fv fragments, diabodies, multivalent binding, linker length, electron microscopy imaging
- MeSH terms
- Animals, Antigen-Antibody Complex, Binding Sites, Immunoglobulin Fragments, Immunoglobulin Idiotypes, Immunoglobulin Fab Fragments, Lymphokines, Microscopy, Electron, Models, Molecular, Protein Conformation, Recombinant Proteins, Sialoglycoproteins, Mice
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