FIELD: chemistry.
SUBSTANCE: invention relates to a method for determination and quantitative estimation of unusually modified glycans, which can be used in analysing glycans. The disclosed method includes the following steps: a) providing a glycan preparation containing unusually modified glycans selected from a group containing sulphated glycans, phosphorylated glycans, polyacetylated sialylated glycans and combinations thereof, where said unusually modified glycans are negatively charged and where said glycan preparation is obtained by separating glycans and sialic acids from a therapeutic glycoprotein composition, where the sialic acids are separated by exposing the therapeutic glycoprotein composition to at least one agent which splits sialic acid residues in conditions which facilitate splitting of sialic acids; b) subjecting the glycan preparation to a chromatographic separation method which separates glycans based on the charge-to-mass ratio, thereby separating said unusually modified glycans; and c) quantitative determination of at least one separated unusually modified glycan using at least one quantitative estimation standard.
EFFECT: novel efficient method which enables to analyse unusually modified glycans.
42 cl, 7 dwg, 2 tbl, 1 ex
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Authors
Dates
2014-08-20—Published
2009-12-18—Filed