Role of Glycosylation in Conformational Stability, Activity, Macromolecular Interaction and Immunogenicity of Recombinant Human Factor VIII
Journal Title: The AAPS Journal - Year 2009, Vol 11, Issue 3
Abstract
Factor VIII (FVIII) is a multi-domain glycoprotein that is an essential cofactor in the blood coagulation cascade. Its deficiency or dysfunction causes hemophilia A, a bleeding disorder. Replacement using exogenous recombinant human factor VIII (rFVIII) is the first line of therapy for hemophilia A. The role of glycosylation on the activity, stability, protein–lipid interaction, and immunogenicity of FVIII is not known. In order to investigate the role of glycosylation, a deglycosylated form of FVIII was generated by enzymatic cleavage of carbohydrate chains. The biochemical properties of fully glycosylated and completely deglycosylated forms of rFVIII (degly rFVIII) were compared using enzyme-linked immunosorbent assay, size exclusion chromatography, and clotting activity studies. The biological activity of degly FVIII decreased in comparison to the fully glycosylated protein. The ability of degly rFVIII to interact with phosphatidylserine containing membranes was partly impaired. Data suggested that glycosylation significantly influences the stability and the biologically relevant macromolecular interactions of FVIII. The effect of glycosylation on immunogenicity was investigated in a murine model of hemophilia A. Studies showed that deletion of glycosylation did not increase immunogenicity.
Authors and Affiliations
Matthew P. Kosloski, Razvan D. Miclea, Sathy V. Balu-Iyer
Keywords
Related Articles
- EP ID EP681482
- DOI 10.1208/s12248-009-9119-y
- Views 84
- Downloads 0
A national survey of U.S. pharmacists in 2000: Assessing nonresponse bias of a survey methodology
The first objective of this study was to assess the existence of nonresponse bias to a national survey of licensed pharmacists conducted in 2000. Three methods were used to assess nonresponse bias. The second objective o...
Current methodology to assess bioequivalence of levothyroxine sodium products is inadequate
Levothyroxine sodium is a drug with a narrow therapeutic index for which an individual patient must have his or her dose carefully titrated to achieve the necessary therapeutic effect. In addition, exogenous levothyroxin...
Preparation, characterization, and biodistribution study of technetium-99m-labeled leuprolide acetate-loaded liposomes in ehrlich ascites tumor-bearing mice
The purpose of this study was to prepare conventional and sterically stabilized liposomes containing leuprolide acetate in an attempt to prolong the biological half life of the drug, to reduce the uptake by reticuloendot...
Mapping the Target Localization and Biodistribution of Non-Radiolabeled VMAT2 Ligands in rat Brain
Imaging targeting vesicular monoamine transporter (VMAT2) alterations is a sensitive tool for early diagnosis of Parkinson’s disease. Our group has reported several novel 2-amino-DTBZ derivatives as potential VMA...
Preparation and in vivo evaluation of SMEDDS (self-microemulsifying drug delivery system) containing fenofibrate
The present work was aimed at formulating a SMEDDS (self-microemulsifying drug delivery system) of fenofibrate and evaluating its in vitro and in vivo potential. The solubility of fenofibrate was determined in various ve...