title: Isoagglutinin reduction by a dedicated immunoaffinity chromatography step in the manufacturing process of human immunoglobulin products.
abstract: BACKGROUND
  The passive transfer of antibodies specific to blood groups A and B (also called isoagglutinins) contained in immunoglobulin (Ig)G products for intravenous administration (IVIG) is believed to be largely responsible for rare but sometimes serious IVIG-related hemolytic events. We present in this work a modification of the manufacturing process of Privigen-a 10% l-proline-stabilized IVIG product-that allows extensive reduction of isoagglutinin concentrations in the final product.
  
  
  METHODS
  An additional immunoaffinity chromatography (IAC) step was introduced toward the end of the manufacturing process of Privigen. Isoagglutinin titers were measured using the indirect agglutination method and a published flow cytometry-based binding assay. Quality attributes, such as microorganism counts and concentration of endotoxins, IgG, IgA, IgM, aggregates, and so forth were measured using standardized procedures.
  
  
  RESULTS
  The introduction of an IAC step in the manufacturing process of Privigen resulted in an 88% to 90% reduction in isoagglutinins between the feed of the chromatography column and the flow-through fraction. All other product quality attributes measured were nearly identical before and after IAC. This process modification resulted in a three-titer-step reduction in isoagglutinin levels in the final IgG product compared to Privigen lots produced by the unmodified process.
  
  
  CONCLUSIONS
  Introducing an isoagglutinin-specific IAC step in the manufacturing process of Privigen is an efficient strategy for reduction of anti-A and anti-B titers. Such reductions might help minimize the risk of hemolytic events in patients receiving IVIG therapy.
authors.affiliation: Research and Development, Berne, Switzerland.
authors.forename: Liane
authors.initials: L
authors.lastname: Hoefferer
authors.affiliation: Process Development, Berne, Switzerland.
authors.forename: Isabelle
authors.initials: I
authors.lastname: Glauser
authors.affiliation: Product Development, Berne, Switzerland.
authors.forename: Annette
authors.initials: A
authors.lastname: Gaida
authors.affiliation: Biochemical Quality Control, Berne, Switzerland.
authors.forename: Katharina
authors.initials: K
authors.lastname: Willimann
authors.affiliation: Biochemical Quality Control, Berne, Switzerland.
authors.forename: Adriano
authors.initials: A
authors.lastname: Marques Antunes
authors.affiliation: Biochemical Quality Control, Berne, Switzerland.
authors.forename: Brigitte
authors.initials: B
authors.lastname: Siani
authors.affiliation: Product Development, Berne, Switzerland.
authors.forename: Sandra
authors.initials: S
authors.lastname: Wymann
authors.affiliation: Biochemical Quality Control, Berne, Switzerland.
authors.forename: Eleonora
authors.initials: E
authors.lastname: Widmer
authors.affiliation: Process Development, Berne, Switzerland.
authors.forename: Ibrahim
authors.initials: I
authors.lastname: El Menyawi
authors.affiliation: Biochemistry Research and Development, Berne, Switzerland.
authors.forename: Reinhard
authors.initials: R
authors.lastname: Bolli
authors.affiliation: Product Development, Berne, Switzerland.
authors.forename: Martin
authors.initials: M
authors.lastname: Spycher
authors.affiliation: Pharmaceutical Development, CSL Behring AG, Berne, Switzerland.
authors.forename: Martin
authors.initials: M
authors.lastname: Imboden
mesh_terms: D001781:Blood Component Removal / Q000379:methods*; D002846:Chromatography, Affinity / Q000379:methods*; D005260:Female; D006388:Hemagglutinins / Q000737:chemistry*; D006801:Humans; D016756:Immunoglobulins, Intravenous / Q000737:chemistry* / Q000302:isolation & purification*; D008297:Male
chemical_list: D006388:Hemagglutinins; D016756:Immunoglobulins, Intravenous