title: Use of purified dermatan sulfate for heparin cofactor II (HC II) assay.
abstract: HC II functional assays are generally preferred with respect to immunochemical assays. Nevertheless functional assays can be biased by the antithrombin III (AT III)-heparin complex activity; in fact trace amounts of heparin generally contaminate dermatan sulfate (DS) commercial preparations used as HC II reaction activators. We have employed a purified DS preparation showing these features: DS concentration 94.4%, chondroitin sulfate 5.6%, M.W. 21.4 kDa. The absence of any interference due to AT III-heparin complexes was verified in a kinetic HC II assay of some human plasma pools. The immunological inhibition of AT III by anti-AT III caused a minimal decrease (6-8%) of the reaction slope, attributable to AT III activity. Progressive increase of heparin concentration in the assay was effective only starting from 30 U/ml (the assay was carried out in the presence of polybrene to prevent any AT III activation). The reference interval (mean +/- SD) obtained from 157 normal subjects was 100.8 +/- 20.2%; there was a good correlation with immunoreactive HC II. The purified DS we have used seems suitable for routinary assays of HC II where a minimal interference due to AT III-heparin is required.
authors.affiliation: Laboratory of Clinical Chemistry, C.O.C., Valeggio, Italy.
authors.forename: O
authors.initials: O
authors.lastname: Ruzzenente
authors.forename: R
authors.initials: R
authors.lastname: Ghidini
authors.forename: G
authors.initials: G
authors.lastname: Poli
authors.forename: R
authors.initials: R
authors.lastname: Schiavon
authors.forename: G C
authors.initials: GC
authors.lastname: Guidi
mesh_terms: D000595:Amino Acid Sequence; D003871:Dermatan Sulfate / Q000302:isolation & purification*; D015844:Heparin Cofactor II / Q000032:analysis*; D006583:Hexadimethrine Bromide; D006801:Humans; D007700:Kinetics; D008969:Molecular Sequence Data
chemical_list: D003871:Dermatan Sulfate; D006583:Hexadimethrine Bromide; D015844:Heparin Cofactor II