title: Fractionation of follicle stimulating hormone charge isoforms in their native form by preparative electrophoresis technology.
abstract: Complex glycoprotein biopharmaceuticals, such as follicle stimulating hormone (FSH), erythropoietin and tissue plasminogen activator consist of a range of charge isoforms due to the extent of sialic acid capping of the glycoprotein glycans. Sialic acid occupies the terminal position on the oligosaccharide chain, masking the penultimate sugar residue, galactose from recognition and uptake by the hepatocyte asialoglycoprotein receptor. It is therefore well established that the more acidic charge isoforms of glycoprotein biopharmaceuticals have higher in vivo potencies than those of less acidic isoforms due to their longer serum half-life. Current strategies for manipulating glycoprotein charge isoform profile involve cell engineering or altering bioprocesss parameters to optimise expression of more acidic or basic isoforms, rather than downstream separation of isoforms. A method for the purification of a discrete range of bioactive recombinant human FSH (rhFSH) charge isoforms based on Gradiflowtrade mark preparative electrophoresis technology is described. Gradiflowtrade mark electrophoresis is scaleable, and incorporation into glycoprotein biopharmaceutical production bioprocesses as a potential final step facilitates the production of biopharmaceutical preparations of improved in vivo potency.
authors.affiliation: Bioengineering Centre, School of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney, Australia.
authors.forename: Dallia
authors.initials: D
authors.lastname: Catzel
authors.forename: David Y
authors.initials: DY
authors.lastname: Chin
authors.forename: Peter G
authors.initials: PG
authors.lastname: Stanton
authors.forename: Peter P
authors.initials: PP
authors.lastname: Gray
authors.forename: Stephen M
authors.initials: SM
authors.lastname: Mahler
mesh_terms: D000818:Animals; D001709:Biotechnology / Q000379:methods*; D016466:CHO Cells; D005591:Chemical Fractionation / Q000379:methods*; D006224:Cricetinae; D003412:Cricetulus; D004586:Electrophoresis / Q000379:methods*; D005640:Follicle Stimulating Hormone / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D015202:Protein Engineering / Q000379:methods*; D020033:Protein Isoforms / Q000235:genetics / Q000302:isolation & purification
chemical_list: D020033:Protein Isoforms; D005640:Follicle Stimulating Hormone