title: Molecular engineering of high affinity single-chain antibody fragment for endothelial targeting of proteins and nanocarriers in rodents and humans.
abstract: Endothelial cells (EC) represent an important target for pharmacologic intervention, given their central role in a wide variety of human pathophysiologic processes. Studies in lab animal species have established that conjugation of drugs and carriers with antibodies directed to surface targets like the Platelet Endothelial Cell Adhesion Molecule-1 (PECAM-1, a highly expressed endothelial transmembrane protein) help to achieve specific therapeutic interventions in ECs. To translate such "vascular immunotargeting" to clinical practice, it is necessary to replace antibodies by advanced ligands that are more amenable to use in humans. We report the molecular design of a single chain variable antibody fragment (scFv) that binds with high affinity to human PECAM-1 and cross-reacts with its counterpart in rats and other animal species, allowing parallel testing in vivo and in human endothelial cells in microfluidic model. Site-specific modification of the scFv allows conjugation of protein cargo and liposomes, enabling their endothelial targeting in these models. This study provides a template for molecular engineering of ligands, enabling studies of drug targeting in animal species and subsequent use in humans.
authors: Department of Pharmacology, Institute for Translational Medicine and Therapeutics, 3400 Civic Center Blvd, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, United States; Center for Targeted Therapeutics and Translational Nanomedicine, Institute for Translational Medicine and Therapeutics, 3400 Civic Center Blvd, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, United States. Electronic address: cgreineder@gmail.com.
authors: Colin F
authors: CF
authors: Greineder
authors: Department of Pharmacology, Institute for Translational Medicine and Therapeutics, 3400 Civic Center Blvd, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, United States; Center for Targeted Therapeutics and Translational Nanomedicine, Institute for Translational Medicine and Therapeutics, 3400 Civic Center Blvd, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, United States.
authors: Elizabeth D
authors: ED
authors: Hood
authors: Department of Pharmacology, Institute for Translational Medicine and Therapeutics, 3400 Civic Center Blvd, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, United States; Center for Targeted Therapeutics and Translational Nanomedicine, Institute for Translational Medicine and Therapeutics, 3400 Civic Center Blvd, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, United States.
authors: Anning
authors: A
authors: Yao
authors: Department of Pharmacology, Institute for Translational Medicine and Therapeutics, 3400 Civic Center Blvd, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, United States; Center for Targeted Therapeutics and Translational Nanomedicine, Institute for Translational Medicine and Therapeutics, 3400 Civic Center Blvd, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, United States.
authors: Makan
authors: M
authors: Khoshnejad
authors: Department of Pharmacology, Institute for Translational Medicine and Therapeutics, 3400 Civic Center Blvd, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, United States; Center for Targeted Therapeutics and Translational Nanomedicine, Institute for Translational Medicine and Therapeutics, 3400 Civic Center Blvd, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, United States.
authors: Jake S
authors: JS
authors: Brenner
authors: Department of Pediatrics, Division of Hematology, The Children's Hospital of Philadelphia, Philadelphia, PA 19104, United States.
authors: Ian H
authors: IH
authors: Johnston
authors: Department of Pediatrics, Division of Hematology, The Children's Hospital of Philadelphia, Philadelphia, PA 19104, United States.
authors: Mortimer
authors: M
authors: Poncz
authors: Department of Molecular, Cellular and Developmental Biology, University of California Santa Barbara, Santa Barbara, CA 93106, United States.
authors: Claudia
authors: C
authors: Gottstein
authors: Department of Pharmacology, Institute for Translational Medicine and Therapeutics, 3400 Civic Center Blvd, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, United States; Center for Targeted Therapeutics and Translational Nanomedicine, Institute for Translational Medicine and Therapeutics, 3400 Civic Center Blvd, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, United States.
authors: Vladimir R
authors: VR
authors: Muzykantov
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000915:Antibody Affinity; D002460:Cell Line; D016503:Drug Delivery Systems; D042783:Endothelial Cells / Q000276:immunology; D004727:Endothelium / Q000276:immunology*; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D008081:Liposomes / Q000008:administration & dosage / Q000737:chemistry / Q000276:immunology* / Q000493:pharmacokinetics; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000276:immunology*; D051381:Rats; D057127:Single-Chain Antibodies / Q000008:administration & dosage / Q000737:chemistry / Q000276:immunology* / Q000493:pharmacokinetics
chemical_list: D008081:Liposomes; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D057127:Single-Chain Antibodies
keywords: Endothelial drug delivery; Immunotargeting; PECAM-1; Single chain variable fragment