Comprehensive optimization of a single-chain variable domain antibody fragment as a targeting ligand for a cytotoxic nanoparticle
| Autor: | Ulrik B. Nielsen, Lihui Xu, Kathy Zhang, Daryl C. Drummond, Tad Kornaga, Alexey Lugovskoy, Neeraj Kohli, Yang Jiao, Melissa Geddie, Dmitri B. Kirpotin, Rachel Rennard |
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| Rok vydání: | 2015 |
| Předmět: |
Antibody-drug conjugate
Antibodies Neoplasm media_common.quotation_subject Immunology Yeast display Mice Drug Delivery Systems Antigen Cell Line Tumor Neoplasms Animals Humans Immunology and Allergy Single-chain variable fragment Cytotoxic T cell Internalization media_common Cytotoxins Chemistry Ligand (biochemistry) Molecular biology Recombinant Proteins Amino Acid Substitution Biophysics Nanoparticles Linker Reports Single-Chain Antibodies |
| Zdroj: | mAbs. 7:42-52 |
| ISSN: | 1942-0870 1942-0862 |
| Popis: | Antibody-targeted nanoparticles have the potential to significantly increase the therapeutic index of cytotoxic anti-cancer therapies by directing them to tumor cells. Using antibodies or their fragments requires careful engineering because multiple parameters, including affinity, internalization rate and stability, all need to be optimized. Here, we present a case study of the iterative engineering of a single chain variable fragment (scFv) for use as a targeting arm of a liposomal cytotoxic nanoparticle. We describe the effect of the orientation of variable domains, the length and composition of the interdomain protein linker that connects VH and VL, and stabilizing mutations in both the framework and complementarity-determining regions (CDRs) on the molecular properties of the scFv. We show that variable domain orientation can alter cross-reactivity to murine antigen while maintaining affinity to the human antigen. We demonstrate that tyrosine residues in the CDRs make diverse contributions to the binding affinity and biophysical properties, and that replacement of non-essential tyrosines can improve the stability and bioactivity of the scFv. Our studies demonstrate that a comprehensive engineering strategy may be required to identify a scFv with optimal characteristics for nanoparticle targeting. |
| Databáze: | OpenAIRE |
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