Antibody-Mediated Neutralization of Perfringolysin O for Intracellular Protein Delivery

• Main Authors: Yang, Nicole Jie Yeon (Contributor), Sklaviadis, Demetra (Contributor), Wittrup, Karl Dane (Contributor), Gui, Dan Yi (Contributor), Vander Heiden, Matthew G. (Contributor), Liu, David V. (Author)
• Other Authors: Massachusetts Institute of Technology. Department of Biological Engineering (Contributor), Massachusetts Institute of Technology. Department of Biology (Contributor), Massachusetts Institute of Technology. Department of Chemical Engineering (Contributor), Koch Institute for Integrative Cancer Research at MIT (Contributor), Liu, David V (Contributor)
• Format: Article
• Language: English
• Published: American Chemical Society (ACS), 2016-12-15T15:27:23Z.
• Subjects: Article
• Online Access: Get fulltext

Perfringolysin O (PFO) is a member of the cholesterol-dependent cytolysin (CDC) family of bacterial pore-forming proteins, which are highly efficient in delivering exogenous proteins to the cytoplasm. However, the indiscriminate and potent cytotoxicity of PFO limits its practical use as an intracellular delivery system. In this study, we describe the design and engineering of a bispecific, neutralizing antibody against PFO, which targets reversibly attenuated PFO to endocytic compartments via receptor-mediated internalization. This PFO-based system efficiently mediated the endosomal release of a co-targeted gelonin construct with high specificity and minimal toxicity in vitro. Consequently, the therapeutic window of PFO was improved by more than 5 orders of magnitude. Our results demonstrating that the activity of pore-forming proteins can be controlled by antibody-mediated neutralization present a novel strategy for utilizing these potent membrane-lytic agents as a safe and effective intracellular delivery vehicle.
Massachusetts Institute of Technology/National Institute of General Medical Sciences Biotechnology Training Program
National Institutes of Health (U.S.) (Grant CA101830)