Biparatopic single-domain antibodies against Axl achieve ultra-high affinity through intramolecular engagement
| Autor: | Greg Hussack, Cunle Wu, Thanh-Dung Nguyen, C. Roger MacKenzie, Mehdi Arbabi-Ghahroudi, Kevin A. Henry, Alma Robert, Toya Nath Baral, Yves Durocher, Maria L. Jaramillo, Shalini Raphael |
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| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
0301 basic medicine
Phage display TAM family medicine.drug_class Recombinant Fusion Proteins Biophysics Antibody Affinity VHH CHO Cells Monoclonal antibody Biochemistry Epitope Receptor tyrosine kinase 03 medical and health sciences 0302 clinical medicine Cricetulus Protein Domains Cell Line Tumor medicine Cytotoxic T cell Animals Humans cancer Molecular Biology single-domain antibody biology Cell Death GAS6 Chemistry Receptor Protein-Tyrosine Kinases Axl Cell Biology Single-Domain Antibodies Cell biology Kinetics nanobody 030104 developmental biology Single-domain antibody HEK293 Cells 030220 oncology & carcinogenesis biology.protein receptor tyrosine kinase Antibody Protein Multimerization Immunoglobulin Heavy Chains Camelids New World Protein Binding |
| Popis: | Overexpression of Axl, a TAM-family receptor tyrosine kinase, plays key roles in the formation, growth, and spread of tumors as well as resistance to targeted therapies and chemotherapies. We identified novel llama VHHs against human Axl using multiple complementary phage display selection strategies and characterized a subset of high-affinity VHHs. The VHHs targeted multiple sites in Ig-like domains 1 and 2 of the Axl extracellular domain, including an immunodominant epitope overlapping the site of Gas6 interaction and two additional non-Gas6 competitive epitopes recognized by murine monoclonal antibodies. Only a subset of VHHs cross-reacted with cynomolgus monkey Axl and none recognized mouse Axl. As fusions to human IgG1 Fc, VHH-Fcs bound Axl+ tumor cell lines and mertansine-loaded VHH-Fcs were cytotoxic in vitro against Axl+ cells in proportion to their binding affinities. Engineered biparatopic VHH-VHH heterodimers bound Axl avidly, and a subset of molecules showed dramatically enhanced association rates indicative of intramolecular binding. These VHHs may have applications as modular elements of biologic drugs such as antibody-drug conjugates. |
| Databáze: | OpenAIRE |
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