Differences in carbohydrate composition of FSH preparations detected with lectin-ELISA systems.
FSH is a glycoprotein containing N-linked carbohydrates which exhibit a variety of forms ranging from mono- to multibranched structures. Variation in glycosylation, particularly the degree of terminal sialylation, determines the half-life of the hormone and hence its in vivo bioactivity. The glycoform content of FSH preparations can differ according to the source (e.g. pituitary, urine), cell line (for rDNA-derived material) and selectivity of purification procedures, and may create difficulties in the preparation and characterization of standards and therapeutic products. In order to develop a simple method to detect changes in glycocomposition, an FSH ELISA was modified by the incorporation of lectins of recognized sugar specificity, and used to examine the terminal sugar composition of ampouled preparations of pituitary, urinary and rDNA-derived FSH. FSH was captured with a specific monoclonal antibody (MAb) and detected with either biotinylated anti-FSH MAb (ELISA) or the sugar-specific lectins (L-ELISA) from Triticum vulgaris (sialic acid, SA), Sambucus nigra (alpha 2,6-linked SA), Maackia amurensis (alpha 2,3-linked SA) or Ricinus communis (free terminal galactose; GAL). Relative estimates of the amounts of terminal SA, its different forms and GAL were derived from the L-ELISA/ELISA data compared with the highly sialylated 1st International Standard for pituitary FSH (IS) 83/575. All the FSH preparations had less SA than the IS with the ratio of alpha 2,3- and alpha 2,6-linked SA varying between preparations. The amounts of alpha 2,6-linked SA relative to the IS were not significantly different in the urinary and pituitary preparations whereas alpha 2,3-linked SA in all preparations was generally less than that of the standard.(ABSTRACT TRUNCATED AT 250 WORDS)
Rafferty
B
B
Division of Endocrinology, National Institute for Biological Standards and Control, South Mimms, Potters Bar, Hertfordshire, UK.
Mower
J A
JA
Ward
H L
HL
Rose
M
M
D002236:Carbohydrate Conformation; D002241:Carbohydrates / Q000032:analysis*; D002367:Ricinus communis; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005640:Follicle Stimulating Hormone / Q000737:chemistry*; D005690:Galactose / Q000032:analysis; D006801:Humans; D037102:Lectins; D010835:Phytohemagglutinins; D037121:Plant Lectins; D010947:Plants, Toxic; D054788:Ribosome Inactivating Proteins; D012794:Sialic Acids / Q000032:analysis; D014909:Wheat Germ Agglutinins
D002241:Carbohydrates; D037102:Lectins; D010835:Phytohemagglutinins; D037121:Plant Lectins; C017707:Ricinus communis agglutinin-1; C050600:Sambucus nigra lectins; D012794:Sialic Acids; D014909:Wheat Germ Agglutinins; C100156:leukoagglutinins, plants; D005640:Follicle Stimulating Hormone; D054788:Ribosome Inactivating Proteins; D005690:Galactose