title: Antibody-mediated neutralization of perfringolysin o for intracellular protein delivery.
abstract: 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.
authors.affiliation: †Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, United States.|⊥Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, United States.
authors.forename: Nicole J
authors.initials: NJ
authors.lastname: Yang
authors.affiliation: †Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, United States.|⊥Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, United States.
authors.forename: David V
authors.initials: DV
authors.lastname: Liu
authors.affiliation: ‡Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, United States.
authors.forename: Demetra
authors.initials: D
authors.lastname: Sklaviadis
authors.affiliation: §Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, United States.|⊥Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, United States.
authors.forename: Dan Y
authors.initials: DY
authors.lastname: Gui
authors.affiliation: §Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, United States.|∥Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, Massachusetts 02115, United States.|⊥Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, United States.
authors.forename: Matthew G
authors.initials: MG
authors.lastname: Vander Heiden
authors.affiliation: †Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, United States.|‡Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, United States.|⊥Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, United States.
authors.forename: K Dane
authors.initials: KD
authors.lastname: Wittrup
mesh_terms: D057134:Antibodies, Neutralizing / Q000737:chemistry*; D001427:Bacterial Toxins / Q000737:chemistry*; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D006460:Hemolysin Proteins / Q000737:chemistry*; D006801:Humans; D008954:Models, Biological; D054353:Perforin / Q000737:chemistry
chemical_list: D057134:Antibodies, Neutralizing; D001427:Bacterial Toxins; D006460:Hemolysin Proteins; D054353:Perforin; C020452:Clostridium perfringens theta-toxin
keywords: cholesterol-dependent cytolysin; endosomal release; intracellular delivery; neutralizing antibody; perfringolysin O; protein delivery