title: Design and chemical synthesis of a homogeneous polymer-modified erythropoiesis protein.
abstract: We report the design and total chemical synthesis of "synthetic erythropoiesis protein" (SEP), a 51-kilodalton protein-polymer construct consisting of a 166-amino-acid polypeptide chain and two covalently attached, branched, and monodisperse polymer moieties that are negatively charged. The ability to control the chemistry allowed us to synthesize a macromolecule of precisely defined covalent structure. SEP was homogeneous as shown by high-resolution analytical techniques, with a mass of 50,825 +/-10 daltons by electrospray mass spectrometry, and with a pI of 5.0. In cell and animal assays for erythropoiesis, SEP displayed potent biological activity and had significantly prolonged duration of action in vivo. These chemical methods are a powerful tool in the rational design of protein constructs with potential therapeutic applications.
authors.affiliation: Gryphon Therapeutics, 250 East Grand Avenue, Suite 90, South San Francisco, CA 94080, USA. Gkochendoerfer@gryphonRX.com
authors.forename: Gerd G
authors.initials: GG
authors.lastname: Kochendoerfer
authors.forename: Shiah-Yun
authors.initials: SY
authors.lastname: Chen
authors.forename: Feng
authors.initials: F
authors.lastname: Mao
authors.forename: Sonya
authors.initials: S
authors.lastname: Cressman
authors.forename: Stacey
authors.initials: S
authors.lastname: Traviglia
authors.forename: Haiyan
authors.initials: H
authors.lastname: Shao
authors.forename: Christie L
authors.initials: CL
authors.lastname: Hunter
authors.forename: Donald W
authors.initials: DW
authors.lastname: Low
authors.forename: E Neil
authors.initials: EN
authors.lastname: Cagle
authors.forename: Maia
authors.initials: M
authors.lastname: Carnevali
authors.forename: Vincent
authors.initials: V
authors.lastname: Gueriguian
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Keogh
authors.forename: Heather
authors.initials: H
authors.lastname: Porter
authors.forename: Stephen M
authors.initials: SM
authors.lastname: Stratton
authors.forename: M Con
authors.initials: MC
authors.lastname: Wiedeke
authors.forename: Jill
authors.initials: J
authors.lastname: Wilken
authors.forename: Jie
authors.initials: J
authors.lastname: Tang
authors.forename: Jay J
authors.initials: JJ
authors.lastname: Levy
authors.forename: Les P
authors.initials: LP
authors.lastname: Miranda
authors.forename: Milan M
authors.initials: MM
authors.lastname: Crnogorac
authors.forename: Suresh
authors.initials: S
authors.lastname: Kalbag
authors.forename: Paolo
authors.initials: P
authors.lastname: Botti
authors.forename: Janice
authors.initials: J
authors.lastname: Schindler-Horvat
authors.forename: Laura
authors.initials: L
authors.lastname: Savatski
authors.forename: John W
authors.initials: JW
authors.lastname: Adamson
authors.forename: Ada
authors.initials: A
authors.lastname: Kung
authors.forename: Stephen B H
authors.initials: SB
authors.lastname: Kent
authors.forename: James A
authors.initials: JA
authors.lastname: Bradburne
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D002942:Circular Dichroism; D015195:Drug Design; D004355:Drug Stability; D004591:Electrophoresis, Polyacrylamide Gel; D004920:Erythropoiesis; D004921:Erythropoietin / Q000737:chemistry / Q000494:pharmacology; D006400:Hematocrit; D006801:Humans; D007526:Isoelectric Point; D051379:Mice; D008969:Molecular Sequence Data; D015394:Molecular Structure; D008970:Molecular Weight; D011108:Polymers / Q000138:chemical synthesis* / Q000737:chemistry* / Q000493:pharmacokinetics / Q000494:pharmacology; D017510:Protein Folding; D011506:Proteins / Q000138:chemical synthesis* / Q000737:chemistry* / Q000493:pharmacokinetics / Q000494:pharmacology; D051381:Rats; D017467:Receptors, Erythropoietin / Q000187:drug effects / Q000378:metabolism; D011994:Recombinant Proteins; D021241:Spectrometry, Mass, Electrospray Ionization
chemical_list: D011108:Polymers; D011506:Proteins; D017467:Receptors, Erythropoietin; D011994:Recombinant Proteins; C472014:synthetic erythropoiesis protein; D004921:Erythropoietin