A panel of synthetic antibodies that selectively recognize and antagonize members of the interferon alpha family

Abstract

The 12 distinct subtypes that comprise the interferon alpha (IFNα) family of cytokines possess anti-viral, anti-proliferative and immunomodulatory activities. They are implicated in the etiology and progression of many diseases, and also used as therapeutic agents for viral and oncologic disorders. However, a deeper understanding of their role in disease is limited by a lack of tools to evaluate single subtypes at the protein level. Antibodies that selectively inhibit single IFNα subtypes could enable interrogation of each protein in biological samples and could be used for characterization and treatment of disease. Using phage-displayed synthetic antibody libraries, we have conducted selections against 12 human IFNα subtypes to explore our ability to obtain fine-specificity antibodies that recognize and antagonize the biological signals induced by a single IFNα subtype. For the first time, we have isolated antibodies that specifically recognize individual IFNα subtypes (IFNα2a/b, IFNα6, IFNα8b and IFNα16) with high affinity that antagonize signaling. Our results show that highly specific antibodies capable of distinguishing between closely related cytokines can be isolated from synthetic libraries and can be used to characterize cytokine abundance and function.

A T cell redirection platform for co-targeting dual antigens on solid tumors

“…25 Notably, the trastuzumab framework has proven to be highly robust for supporting many diverse paratopes, and we and others have used this framework to construct naïve phage-displayed libraries that have yielded thousands of Fabs recognizing hundreds of diverse antigens. 29,42,[56][57][58][59] Consequently, virtually any stable Fab derived from natural IgGs or synthetic libraries could be converted into the DATE format to provide a vast toolkit for exploring novel TAAs for T-cell recruitment against diverse tumors. To further enhance specificity in a modular manner, we developed d-DATEs by adding a second tumor-targeting moiety in the form of an autonomous VH domain, built on a modified version of the trastuzumab framework.…”

Antibody selection using clonal cocultivation of Escherichia coli and eukaryotic cells in miniecosystems

“…Phage display is a popular tool for the development of therapeutic antibodies in the pharmaceutical industry. In most cases, therapeutic antibodies have biological functions; for example, antibodies bind to target proteins and act as antagonists, allosteric regulators, or agonists (10,11). The main challenge with the use of phage panning for drug discovery is that the panning is based purely on the affinity between the antibody and the antigen (SI Appendix, Fig.…”