title: Enhancement of therapeutic protein in vivo activities through glycoengineering.
abstract: Delivery of protein therapeutics often requires frequent injections because of low activity or rapid clearance, thereby placing a burden on patients and caregivers. Using glycoengineering, we have increased and prolonged the activity of proteins, thus allowing reduced frequency of administration. Glycosylation analogs with new N-linked glycosylation consensus sequences introduced into the protein were screened for the presence of additional N-linked carbohydrates and retention of in vitro activity. Suitable consensus sequences were combined in one molecule, resulting in glycosylation analogs of rHuEPO, leptin, and Mpl ligand. All three molecules had substantially increased in vivo activity and prolonged duration of action. Because these proteins were of three different classes (rHuEPO is an N-linked glycoprotein, Mpl ligand an O-linked glycoprotein, and leptin contains no carbohydrate), glycoengineering may be generally applicable as a strategy for increasing the in vivo activity and duration of action of proteins. This strategy has been validated clinically for glycoengineered rHuEPO (darbopoetin alfa).
authors.affiliation: Amgen, One Amgen Center, Thousand Oaks, CA 91320, USA. selliott@amgen.com
authors.forename: Steve
authors.initials: S
authors.lastname: Elliott
authors.forename: Tony
authors.initials: T
authors.lastname: Lorenzini
authors.forename: Sheilah
authors.initials: S
authors.lastname: Asher
authors.forename: Ken
authors.initials: K
authors.lastname: Aoki
authors.forename: David
authors.initials: D
authors.lastname: Brankow
authors.forename: Lynette
authors.initials: L
authors.lastname: Buck
authors.forename: Leigh
authors.initials: L
authors.lastname: Busse
authors.forename: David
authors.initials: D
authors.lastname: Chang
authors.forename: Janis
authors.initials: J
authors.lastname: Fuller
authors.forename: James
authors.initials: J
authors.lastname: Grant
authors.forename: Natasha
authors.initials: N
authors.lastname: Hernday
authors.forename: Martha
authors.initials: M
authors.lastname: Hokum
authors.forename: Sylvia
authors.initials: S
authors.lastname: Hu
authors.forename: Andrew
authors.initials: A
authors.lastname: Knudten
authors.forename: Nancy
authors.initials: N
authors.lastname: Levin
authors.forename: Renee
authors.initials: R
authors.lastname: Komorowski
authors.forename: Frank
authors.initials: F
authors.lastname: Martin
authors.forename: Rachell
authors.initials: R
authors.lastname: Navarro
authors.forename: Timothy
authors.initials: T
authors.lastname: Osslund
authors.forename: Gary
authors.initials: G
authors.lastname: Rogers
authors.forename: Norma
authors.initials: N
authors.lastname: Rogers
authors.forename: Geri
authors.initials: G
authors.lastname: Trail
authors.forename: Joan
authors.initials: J
authors.lastname: Egrie
mesh_terms: D000740:Anemia / Q000188:drug therapy; D000818:Animals; D016466:CHO Cells / Q000378:metabolism; D019556:COS Cells / Q000378:metabolism; D006224:Cricetinae; D016503:Drug Delivery Systems / Q000379:methods*; D004921:Erythropoietin / Q000235:genetics / Q000378:metabolism / Q000627:therapeutic use; D005079:Excipients / Q000737:chemistry; D005260:Female; D024861:Genetic Enhancement / Q000379:methods; D006023:Glycoproteins / Q000096:biosynthesis* / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D020738:Leptin / Q000096:biosynthesis / Q000172:deficiency / Q000235:genetics / Q000627:therapeutic use; D051379:Mice; D008807:Mice, Inbred BALB C; D015202:Protein Engineering / Q000379:methods*; D011506:Proteins / Q000008:administration & dosage / Q000627:therapeutic use*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000627:therapeutic use; D013926:Thrombopoietin / Q000096:biosynthesis / Q000627:therapeutic use
chemical_list: D005079:Excipients; D006023:Glycoproteins; D020738:Leptin; D011506:Proteins; D011994:Recombinant Proteins; D004921:Erythropoietin; D013926:Thrombopoietin