Internal carbohydrate complexity of the oligosaccharide chains of recombinant human follicle stimulating hormone (Puregon, Org 32489): a comparison with Metrodin and Metrodin-HP.
Glycoforms of recombinant human follicle stimulating hormone (rhFSH) (Org 32489, Puregon) were characterized using concanavalin A lectin affinity chromatography to reveal information about the internal carbohydrate complexity (extent of carbohydrate side-chain branching) of the preparations. The rhFSH glycoforms were measured by radioimmunoassay and a two-site immunoradiometric assay and compared with those in two urinary preparations (Metrodin and Metrodin-HP) used in assisted reproduction programmes and a urinary FSH international standard 70/45 (uFSH IS 70/45). Similar data were obtained with both assays; rhFSH had 6% complex internal carbohydrate structures compared with 22-27% for Metrodin, Metrodin-HP and uFSH. The proportion of simple carbohydrate structures was also different, with rhFSH having 18.5 compared with 4.5-9.3% for Metrodin, Metrodin-HP and uFSH. A linear relationship was observed between the percentage glycoforms with an isoelectric point (pl) < 4 and the log percentage simple forms (logarithmic regression; r = 0.93) indicating a direct relationship between carbohydrate complexity and charge heterogeneity. In summary, rhFSH contains fewer complex forms and an increased proportion of simple carbohydrate structures in comparison with Metrodin, Metrodin-HP and IS 70/45.
Harris
S D
SD
Department of Medicine, University of Manchester, Hope Hospital, Salford, UK.
Anobile
C J
CJ
McLoughlin
J D
JD
Mitchell
R
R
Lambert
A
A
Robertson
W R
WR
D002241:Carbohydrates / Q000032:analysis*; D002846:Chromatography, Affinity; D003208:Concanavalin A; D005640:Follicle Stimulating Hormone / Q000737:chemistry* / Q000652:urine; D043373:Follicle Stimulating Hormone, Human; D006031:Glycosylation; D006801:Humans; D011863:Radioimmunoassay; D011994:Recombinant Proteins / Q000737:chemistry; D012015:Reference Standards
D002241:Carbohydrates; D043373:Follicle Stimulating Hormone, Human; D011994:Recombinant Proteins; D003208:Concanavalin A; D005640:Follicle Stimulating Hormone