Variable Domains

1995 ◽  
pp. 193-247 ◽  
Author(s):  
Herbert Amann
Keyword(s):  
Variable Domains

scholarly journals Site-Specific Dual-Labeling of a VHH with a Chelator and a Photosensitizer for Nuclear Imaging and Targeted Photodynamic Therapy of EGFR-Positive Tumors

Cancers ◽  
2021 ◽  
Vol 13 (3) ◽  
pp. 428
Author(s):  
Emma Renard ◽  
Estel Collado Camps ◽  
Coline Canovas ◽  
Annemarie Kip ◽  
Martin Gotthardt ◽  
...  
Keyword(s):  
Photodynamic Therapy ◽  
Fluorescence Imaging ◽  
Ex Vivo ◽  
Growth Factor Receptor ◽  
Nuclear Imaging ◽  
A431 Cells ◽  
Site Specific ◽  
Variable Domains ◽  
Diagnostic Probe

Variable domains of heavy chain only antibodies (VHHs) are valuable agents for application in tumor theranostics upon conjugation to both a diagnostic probe and a therapeutic compound. Here, we optimized site-specific conjugation of the chelator DTPA and the photosensitizer IRDye700DX to anti-epidermal growth factor receptor (EGFR) VHH 7D12, for applications in nuclear imaging and photodynamic therapy. 7D12 was site-specifically equipped with bimodal probe DTPA-tetrazine-IRDye700DX using the dichlorotetrazine conjugation platform. Binding, internalization and light-induced toxicity of DTPA-IRDye700DX-7D12 were determined using EGFR-overexpressing A431 cells. Finally, ex vivo biodistribution of DTPA-IRDye700DX-7D12 in A431 tumor-bearing mice was performed, and tumor homing was visualized with SPECT and fluorescence imaging. DTPA-IRDye700DX-7D12 was retrieved with a protein recovery of 43%, and a degree of labeling of 0.56. Spectral properties of the IRDye700DX were retained upon conjugation. 111In-labeled DTPA-IRDye700DX-7D12 bound specifically to A431 cells, and they were effectively killed upon illumination. DTPA-IRDye700DX-7D12 homed to A431 xenografts in vivo, and this could be visualized with both SPECT and fluorescence imaging. In conclusion, the dichlorotetrazine platform offers a feasible method for site-specific dual-labeling of VHH 7D12, retaining binding affinity and therapeutic efficacy. The flexibility of the described approach makes it easy to vary the nature of the probes for other combinations of diagnostic and therapeutic compounds.

RT-PCR cloning and characterization of mouse immunoglobulin variable domains with high affinity for HLA-DR antigens

1997 ◽  
Vol 46 (3) ◽  
pp. 249-250 ◽  
Author(s):  
V. P. Eswarakumar ◽  
M. C. Raja ◽  
V. R. Muthukkaruppan
Keyword(s):  
Rt Pcr ◽  
Pcr Cloning ◽  
High Affinity ◽  
Hla Dr ◽  
Mouse Immunoglobulin ◽  
Variable Domains

scholarly journals Antibody light chain variable domains and their biophysically improved versions for human immunotherapy

mAbs ◽  
2013 ◽  
Vol 6 (1) ◽  
pp. 219-235 ◽  
Author(s):  
Dae Young Kim ◽  
Rebecca To ◽  
Hiba Kandalaft ◽  
Wen Ding ◽  
Henk van Faassen ◽  
...  
Keyword(s):  
Light Chain ◽  
Variable Domains

scholarly journals A Two-Step Approach for the Design and Generation of Nanobodies

2018 ◽  
Vol 19 (11) ◽  
pp. 3444 ◽  
Author(s):  
Hanna Wagner ◽  
Sarah Wehrle ◽  
Etienne Weiss ◽  
Marco Cavallari ◽  
Wilfried Weber
Keyword(s):  
Heavy Chain ◽  
Animal Experiments ◽  
Affinity Maturation ◽  
Phage Library ◽  
Second Step ◽  
Selection Procedures ◽  
Variable Domains ◽  
Phage Libraries ◽  
Complementarity Determining Regions ◽  
Conventional Antibody

Nanobodies, the smallest possible antibody format, have become of considerable interest for biotechnological and immunotherapeutic applications. They show excellent robustness, are non-immunogenic in humans, and can easily be engineered and produced in prokaryotic hosts. Traditionally, nanobodies are selected from camelid immune libraries involving the maintenance and treatment of animals. Recent advances have involved the generation of nanobodies from naïve or synthetic libraries. However, such approaches demand large library sizes and sophisticated selection procedures. Here, we propose an alternative, two-step approach for the design and generation of nanobodies. In a first step, complementarity-determining regions (CDRs) are grafted from conventional antibody formats onto nanobody frameworks, generating weak antigen binders. In a second step, the weak binders serve as templates to design focused synthetic phage libraries for affinity maturation. We validated this approach by grafting toxin- and hapten-specific CDRs onto frameworks derived from variable domains of camelid heavy-chain-only antibodies (VHH). We then affinity matured the hapten binder via panning of a synthetic phage library. We suggest that this strategy can complement existing immune, naïve, and synthetic library based methods, requiring neither animal experiments, nor large libraries, nor sophisticated selection protocols.

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