title: The conformation of the poly(ethylene glycol) chain in mono-PEGylated lysozyme and mono-PEGylated human growth hormone.
abstract: Covalent conjugation of poly(ethylene glycol) or "PEGylation" has proven an effective strategy to improve pharmaceutical protein efficacy by hindering recognition by proteases, inhibitors, and antibodies and by retarding renal clearance. Because it determines the strength and range of intermolecular steric forces and the hydrodynamic properties of the conjugates, the configuration of protein-conjugated PEG chains is the key factor determining how PEGylation alters protein in vivo circulation time. Mono-PEGylated proteins are typically described as having a protective PEG shroud wrapped around the protein, but recent dynamic light scattering studies suggested that conjugates adopt a dumbbell configuration, with a relatively unperturbed PEG random coil adjacent to the globular protein. We used small-angle neutron scattering (SANS) to distinguish between the dumbbell model and the shroud model for chicken-egg lysozyme and human growth hormone covalently conjugated to a single 20 kDa PEG chain. The SANS contrast variation technique was used to isolate the PEG portion of the conjugate. Scattering intensity profiles were well described by the dumbbell model and inconsistent with the shroud model.
authors.affiliation: Center for Complex Fluids Engineering, Department of Chemical Engineering, Carnegie Mellon University , Pittsburgh, PA 15213, USA.
authors.forename: Sheetal S
authors.initials: SS
authors.lastname: Pai
authors.forename: Boualem
authors.initials: B
authors.lastname: Hammouda
authors.forename: Kunlun
authors.initials: K
authors.lastname: Hong
authors.forename: Danilo C
authors.initials: DC
authors.lastname: Pozzo
authors.forename: Todd M
authors.initials: TM
authors.lastname: Przybycien
authors.forename: Robert D
authors.initials: RD
authors.lastname: Tilton
mesh_terms: D000818:Animals; D002645:Chickens; D019382:Human Growth Hormone / Q000737:chemistry*; D006801:Humans; D008958:Models, Molecular; D008968:Molecular Conformation; D009113:Muramidase / Q000737:chemistry*; D011092:Polyethylene Glycols / Q000737:chemistry*
chemical_list: D019382:Human Growth Hormone; D011092:Polyethylene Glycols; D009113:Muramidase