title: Pharmacokinetic in vivo comparison using 1-stage and chromogenic substrate assays with two formulations of Hemofil-M.
abstract: In a study to demonstrate the safety and pharmacokinetics (half-life and recovery) of two different method M purified AHF (Hemofil-M) concentrates processed in the USA and Spain, two different methods of factor VIII assay (one-stage clotting and chromogenic) have been compared in vivo. The study was a single centre blinded, randomised, crossover study. Twelve patients with severe haemophilia A (VIII:C < 2 u/dl) were divided into two subgroups of six. None had received factor VIII concentrate within 48 h preceding the study. Twenty-four pharmacokinetic studies were performed in the 12 patients. Each subgroup received two different lots of study material (US and Spanish) at a dose of 50 u/kg seven days apart. A second randomisation was nominal potency, high: 1000 u or mid: 500 u per vial. The potency label was a one-stage clotting assay using the mega I standard. A standard pharmacokinetic study was performed over 24 h and each blinded sample was analysed in duplicate by a one-stage clotting (aPTT) and a chromogenic (Chromogenix AB; CS) assay at the Royal Free and NIBSC. Pharmacokinetic modelling was performed. The mean label for Hemofil-M using the chromogenic substrate assay was 79% that using the one stage assay (Mega I standard). The recovery was 17-28% higher measured by chromogenic compared to the clotting assay. Since most clinicians use the clotting assay, potency labelling using the chromogenic assay, will overestimate predicted Hemofil-M recovery by as much as 25%.
authors.affiliation: Haemophilia Centre, Royal Free Hospital, London, UK.
authors.forename: C
authors.initials: C
authors.lastname: Lee
authors.forename: T
authors.initials: T
authors.lastname: Barrowcliffe
authors.forename: G
authors.initials: G
authors.lastname: Bray
authors.forename: E
authors.initials: E
authors.lastname: Gomperts
authors.forename: A
authors.initials: A
authors.lastname: Hubbard
authors.forename: G
authors.initials: G
authors.lastname: Kemball-Cook
authors.forename: P
authors.initials: P
authors.lastname: Lilley
authors.forename: D
authors.initials: D
authors.lastname: Owens
authors.forename: L
authors.initials: L
authors.lastname: Von Tilberg
authors.forename: J
authors.initials: J
authors.lastname: Pasi
mesh_terms: D001681:Biological Assay / Q000379:methods*; D005169:Factor VIII / Q000008:administration & dosage / Q000493:pharmacokinetics*; D006207:Half-Life; D006801:Humans; D013379:Substrate Specificity
chemical_list: D005169:Factor VIII