title: Molecular modifications in factor VIII concentrates produced from different plasma pools.
abstract: We defined the main cause for the increased immunogenicity of the commercial factor VIII (fVIII) plasma-derived concentrates reported to induce formation of inhibitory antibodies in haemophilia A patients in Germany and Belgium. Formation of these antibodies directed against the C2 domain of fVIII was previously attributed to the use of solvent/detergent (S/D) treatment and pasteurisation for virus inactivation of fVIII concentrate. Since fVIII concentrates associated with increased immunogenicity were prepared from plasma pools characterised by elevated levels of coagulation markers, we examined whether the plasma source or S/D treatment and pasteurisation are responsible for structural changes within the C2 domain causing its abnormal immunogenicity. We found that samples of fVIII concentrate that originated from the abnormal plasma pool had a reduced ability to bind to phospholipid and conformationally sensitive anti-C2 domain antibodies, this effect being mostly pronounced in the samples that underwent both S/D treatment and pasteurisation. Thus, our study suggests that insufficient quality of the starting plasma pools is the major factor determining the structural alterations in the C2 domain of fVIII, whereas combination of S/D treatment and pasteurisation aggravates these changes.
authors.affiliation: Department of Biochemistry, Holland Laboratory, American Red Cross, 15601 Crabbs Branch Way, Rockville, MD 20855, USA. saenko@usa.redcross.org
authors.forename: E
authors.initials: E
authors.lastname: Saenko
authors.forename: D
authors.initials: D
authors.lastname: Josic
authors.forename: M
authors.initials: M
authors.lastname: Stadler
authors.forename: A
authors.initials: A
authors.lastname: Sarafanov
authors.forename: Y
authors.initials: Y
authors.lastname: Lim
authors.forename: M
authors.initials: M
authors.lastname: Shima
authors.forename: N
authors.initials: N
authors.lastname: Ananyeva
authors.forename: H
authors.initials: H
authors.lastname: Schwinn
mesh_terms: D000990:Antithrombin III / Q000378:metabolism; D003902:Detergents / Q000494:pharmacology; D005169:Factor VIII / Q000737:chemistry* / Q000187:drug effects / Q000276:immunology; D005340:Fibrinogen / Q000378:metabolism; D006467:Hemophilia A / Q000150:complications / Q000188:drug therapy; D006801:Humans; D010447:Peptide Hydrolases / Q000378:metabolism; D010743:Phospholipids / Q000378:metabolism; D010949:Plasma / Q000737:chemistry; D011485:Protein Binding / Q000187:drug effects; D017434:Protein Structure, Tertiary / Q000187:drug effects; D012997:Solvents / Q000494:pharmacology; D013242:Sterilization
chemical_list: D003902:Detergents; D010743:Phospholipids; D012997:Solvents; C046193:antithrombin III-protease complex; D000990:Antithrombin III; D005169:Factor VIII; D005340:Fibrinogen; D010447:Peptide Hydrolases