title: Reduction of Isoagglutinin in Intravenous Immunoglobulin (IVIG) Using Blood Group A- and B-Specific Immunoaffinity Chromatography: Industry-Scale Assessment.
abstract: BACKGROUND
  Hemolysis, a rare but potentially serious complication of intravenous immunoglobulin (IVIG) therapy, is associated with the presence of antibodies to blood groups A and B (isoagglutinins) in the IVIG product. An immunoaffinity chromatography (IAC) step in the production process could decrease isoagglutinin levels in IVIG.
  
  
  OBJECTIVE
  Our objectives were to compare isoagglutinin levels in a large number of IVIG (Privigen®) batches produced with or without IAC and to assess the feasibility of the production process with an IAC step on an industrial scale.
  
  
  METHODS
  The IAC column comprised a blend of anti-A and anti-B resins formed by coupling synthetic blood group antigens (A/B-trisaccharides) to a base bead matrix, and was introduced towards the end of the industrial-scale IVIG manufacturing process. Isoagglutinin levels in IVIG were determined by anti-A and anti-B hemagglutinin direct and indirect methods according to the European Pharmacopoeia (Ph. Eur.) and an isoagglutinin flow cytometry assay. IVIG product quality was assessed with respect to the retention of immunoglobulin G (IgG) subclasses, specific antibodies, and removal of IgM using standardized procedures.
  
  
  RESULTS
  The IAC step reduced isoagglutinins in IVIG by two to three titer steps compared with lots produced without IAC. The median anti-A and anti-B titers with IAC were 1:8 and 1:4, respectively, when measured by the Ph. Eur. direct method, and 1:2 and <1, respectively, when measured by the Ph. Eur. indirect method. The isoagglutinin flow cytometry assay showed an 87-90 % reduction in isoagglutinins in post-IAC versus pre-IAC fractions. IAC alone reduced anti-A and anti-B of the IgMs isotype by 92.5-97.8 % and 95.4-99.2 %, respectively. Other product quality characteristics were similar with and without IAC.
  
  
  CONCLUSIONS
  IAC is an effective method for reducing isoagglutinin levels in IVIG, and it is feasible on an industrial scale.
authors.affiliation: Department of Research and Development, CSL Behring AG, Wankdorfstrasse 10, 3000, Bern 22, Switzerland. simon.gerber@cslbehring.com.
authors.forename: Simon
authors.initials: S
authors.lastname: Gerber
authors.affiliation: Department of Research and Development, CSL Behring AG, Wankdorfstrasse 10, 3000, Bern 22, Switzerland.
authors.forename: Annette
authors.initials: A
authors.lastname: Gaida
authors.affiliation: Department of Quality Control, CSL Behring AG, Bern, Switzerland.
authors.forename: Nicole
authors.initials: N
authors.lastname: Spiegl
authors.affiliation: Department of Research and Development, CSL Behring AG, Wankdorfstrasse 10, 3000, Bern 22, Switzerland.
authors.forename: Sandra
authors.initials: S
authors.lastname: Wymann
authors.affiliation: Department of Quality Control, CSL Behring AG, Bern, Switzerland.
authors.forename: Adriano Marques
authors.initials: AM
authors.lastname: Antunes
authors.affiliation: Department of Research and Development, CSL Behring AG, Wankdorfstrasse 10, 3000, Bern 22, Switzerland.
authors.forename: Ibrahim El
authors.initials: IE
authors.lastname: Menyawi
authors.affiliation: Department of Project Management, CSL Behring AG, Bern, Switzerland.
authors.forename: Brigitte
authors.initials: B
authors.lastname: Zurbriggen
authors.affiliation: Department of Medical Affairs, CSL Behring AG, Bern, Switzerland.
authors.forename: Alphonse
authors.initials: A
authors.lastname: Hubsch
authors.affiliation: Department of Research and Development, CSL Behring AG, Wankdorfstrasse 10, 3000, Bern 22, Switzerland.
authors.forename: Martin
authors.initials: M
authors.lastname: Imboden
mesh_terms: D000017:ABO Blood-Group System / Q000097:blood; D002846:Chromatography, Affinity / Q000379:methods*; D046148:Donor Selection; D004345:Drug Industry / Q000379:methods; D005434:Flow Cytometry / Q000379:methods; D006388:Hemagglutinins / Q000097:blood*; D006801:Humans; D007074:Immunoglobulin G; D016756:Immunoglobulins, Intravenous / Q000302:isolation & purification*; D011786:Quality Control
chemical_list: D000017:ABO Blood-Group System; D006388:Hemagglutinins; D007074:Immunoglobulin G; D016756:Immunoglobulins, Intravenous