Antihemophilic Factor (Human), Method M, Monoclonal Purified - Hemofil M; Factor VIII:von Wille-brand’s factor complex; Factor VIII:C
Status: approved; marketed
Organizations involved:
Baxter Hyland Immuno – Manuf.; R&D; Tech.; World mark.
Baxter Healthcare Corp. – Parent
Genetics Institute, Inc. – Tech.; Former
Wyeth – Parent; Former
New York Blood Center, Inc. – Tech.
Cross ref: See the entries for Factor VIII Products (#715) and for other monoclonal antibody purified Factor VIII products. See also a nearly identical product, Monarc-M (#721), also manufactured by Baxter Hyland Immuno, formerly marketed by the American Red Cross, now marketed by Baxter Hyland Immuno.
Description: Antihemophilic Factor (Human), Method M, Monoclonal Purified or Hemophil M is a lyophilized (freeze-dried) formulation of a Factor VIII:C (Factor VIII:von Wille-brand’s factor complex; FVIII:vWF) manufactured from pooled Plasma with processing including immunoaffinity chromatography (Method M using matrix-bound murine Factor VIII monoclonal antibodies), ion exchange chromatography, and viral inactivation using the solvent detergent process which substantially reduces titers of enveloped viruses. Monarc-M is a highly purified Factor VIII:C product. It has an activity range of 2 to 15 International Units/mg of total protein; with a specific activity of approximately 2,000 International Units/mg of protein before addition of Albumin (Human) as a stabilizer. The protein in greatest concentration in Monarc-M is Albumin (Human).
Monarc-M, formerly manufactured for the American Red Cross (ARC) by Baxter Hyland Immuno (now marketed by Baxter Hyland Immuno), is the same as Hemofil M manufactured and marketed by Baxter Hyland Immuno, except the Plasma and Albumin (Human) used as stabilizer are obtained from ARC volunteer donors, while Hemophil M is manufactured from Plasma and Albumin (Human) obtained primarily from paid plasmapheresis donors.
Each bottle of Hemofil M is labeled with its Factor VIII activity content expressed in International Units per bottle, which is referenced to the WHO International Standard. After reconstitution with the appropriate volume of diluent, the product contains approximately 12.5 mg/mL Albumin (Human), 1.5 mg/mL polyethylene glycol (3350), 0.055 M histidine and 0.030 M glycine as stabilizing agents. Hemofil M also contains, per AHF International Unit, not more than 0.1 ng mouse protein (from immunoaffinity purification), and residues from solvent detergent viral inactivation – 18 ng organic solvent (tri-n-butyl phosphate) and 50 ng detergent (octoxynol; Triton X-100).
Nomenclature: Factor VIII:vWF, Mab purif./Baxter [BIO]; Hemofil M [TR]; Antihemophilic Factor (Human), Method M, Monoclonal Purified [FDA used on insert/labeling]; Method-M [TR for immunoaffinity purification process]; Antihemophilic Factor (Human) [FDA]; Antihemophilic Factor [USAN]; Antihemophilic Factor, Human [USAN former]; Antihemophilic Factor USP [USP]; Blood-coagulation factor VIII, complex [CAS]; 9001-27-8 [CAS RN]; Factor VIII:von Willebrand’s Factor complex [SY]; Antihemophilic Factor VIII [SY]; Haemophil, Method M [TR foreign]; FVIII:vWF [SY]; AHF [SY]
Companies.: Hemofil M was developed and is manufactured by Baxter Hyland Immuno (Glendale, CA), CBER/FDA est. no. 0140, a subsidiary of Baxter Healthcare Corp. Hemophil M is marketed in the U.S. by Baxter Hyland Immuno and internationally by Baxter affiliates.
Manufacture: manufacture starts with Cryoprecipitated AHF (#716). Cryoprecipitation provides the fraction of frozen pooled human plasma rich in Factor VIII and other coagulation factors. Source material (e.g., Cryoprecipitated AHF) may be provided from other U.S. licensed manufacturers. The cryoprecipitate is put through the 3-step “Method M” process:
a) solvent detergent viral inactivation - The cryoprecipitate is mixed with solvent (tri-n-butyl phosphate) and detergent (Triton X-100 from Rohm & Haas; octoxynol; polyethylene glycol p-isooctylphenyl ether). This is highly effective for inactivating lipid enveloped viruses, e.g., HIV, hepatitis B virus (HBV), and hepatitis C virus (HCV).
b) immunoaffinity chromatography - A column with a matrix of immobilized Factor VIII:C monoclonal antibodies is used to selectively adsorb and then elute (release) Factor VIII. The process uses F8.1 monoclonal antibody produced by hybridoma GI-F8/1.5.6 (ATCC HB 11552) developed by Genetics Institute, Inc., now Wyeth, with specificity for the 90 kDa heavy chain of Factor VIII (not von Willebrand’s factor). The same F8.1 monoclonal antibody is used for purification of Baxter’s two recombinant Factor VIII products, Recombinate and Advate.
c) ion-exchange chromatography - Q-Sepharose column chromatography further purifies Factor VIII. This process also further partitions and removes viruses and extraneous proteins.
Method M processing substantially removes non-Factor VIII proteins, lipid enveloped viruses, non-lipid marker viruses (e.g., reovirus and encephalomyocarditis virus), and residual solvent and detergent.
Method M processing substantially reduces titers of lipid enveloped viruses, e.g., HIV-1 by 10.3 log (1010.3), bovine diarrhea virus (BDV) by 6.1 log, porcine respiratory virus (PRV) by 5.4 log; and non-lipid enveloped viruses, e.g., porcine parvovirus (PPV) by 5.6 log and hepatitis A virus (HAV) by 7.2 log. When Sindbis virus, vesicular stomatitis virus (VSV), and pseudorabies virus were added during manufacture, Method M was shown to inactivate 3-4 logs of these viruses. The infectivity of HIV seeded into cryoprecipitate was reduced by greater than 4 logs almost instantaneously by the organic solvent/detergent step. Besides viral inactivation (killing) by the solvent detergent viral inactivation process, the immunoaffinity purification partitions out and removes substantial amounts of viral contaminants. Hepatitis A Virus (HAV; pHM-A5, clone 24A) removal studies were conducted during manufacture using scaled-down, validated immunoaffinity chromatography columns. Samples collected during the process were assayed for HAV using fetal rhesus monkey kidney cells (FRhMk-4) in a TCID50 assay. In three experiments, the concentration of HAV was decreased by >4.7 log. In four other experiments, the concentrations of both enveloped and non-enveloped viruses were decreased by approximately 4 logs during the immunoaffinity chromatography step.
Note, a heat treatment (pasteurization) step was added for viral inactivation in 1983, shortly after HIV and its potential for contamination of blood products became known. This is no longer used during manufacture of Hemofil M (and Monarc-M). The Method M viral inactivation/removal steps are more effective than heat treatment alone.
See also the Tech. transfer section for descriptions of manufacturing technologies.
FDA class: Biologic PLA
CBER class: Blood And Blood Derivatives
Approvals: Date = 19660311; first approval, PLA, for Antihemophilic Factor (Human) granted to Baxter
Date = 19830321; PLA supplement; Indication = addition of a heat treatment viral inactivation process during manufacture
Date = 19960311; PLA supplement; Indication = addition of immunoaffinity purification (first approval of monoclonal purified product, Hemofil M)
Indications: [full text of "INDICATIONS AND USAGE” section from recent product insert/labeling]:
The use of Hemofil M is indicated in hemophilia A (classical hemophilia) for the prevention and control of hemorrhagic episodes. Hemofil M can be of significant therapeutic value in patients with acquired Factor VIII inhibitors not exceeding 10 Bethesda Units per mL. However, in such uses, the dosage should be controlled by frequent laboratory determinations of circulating AHF.
Hemofil M is not indicated in von Willebrand’s disease.
Tech. transfer: U.S. patent 5,470,954, Neslund, et al, “Ultra-purification Process for Factor VIII,” Nov. 28, 1995, assigned to Baxter International Inc., describes “a process for purifying Factor VIII:C comprising contacting an immobilized antibody specifically binding a Factor VIII:C with Factor VIII: C, desorbing Factor VIII:C from the antibody which had adsorbed it, eluting Factor VIII:C from the presence of the antibody, passing the eluted Factor VIII:C through an affinity region capable of binding the Factor VIII:C, binding the Factor VIII:C in the affinity region and passing contaminants through said region, and eluting the purified Factor VIII:C.”
The monoclonal antibody used for Method M immunoaffinity chromatography purification was licensed from Gene-tics Institute, Inc., then a subsidiary of American Home Products Corp., now Wyeth. The hybridoma GI-F8/1.5.6 was deposited with the American Type Culture Collection as ATCC HB 11552. This hybridoma cell line was prepared by conventional murine hybridoma technology (method of Milstein and Kohler). BALB/c female mice were immunized with human Factor VIII:C, the spleen cells of the immunized mice were fused with P-3Ag 8653 murine myeloma cells, and the resultant hybridomas were screened for supernatants containing monoclonal antibody with selective binding to human Factor VIII. The desired hybridoma was cloned and characterized. Hybridoma GI-F8/1.5.6 expresses a monoclonal antibody against an epitope on human Factor VIII protein and binds plasma-derived and recombinant Factor VIII.
Solvent detergent viral inactivation technology was developed by and nonexclusively licensed from the New York Blood Center, e.g., U.S. patent 4,820,805. See the entry for Pooled Plasma, Solvent Detergent Treated (SD Plasma; #799) for further information about solvent detergent viral inactivation, used primarily for inactivation of enveloped viruses (e.g., HIV, hepatitis B and C viruses).
Market: The 2007 Average Wholesale Price (AWP) is $1.07/IU irrespective of vial size (Red Book, 2007).
Medicare reimbursement is set at $0.90/IU for inpatient and home care, and $0.51/IU for outpatient care. The Estimated Acquisition Costs (for hospitals, treatment centers) is $0.60-$0.66/IU [from NHF].
In its March, 2006 price list, FFF Enterprises, a major biologics distributor, reported its price as $0.71/IU for all sizes (same in 2005).
Companies involvement:
Full monograph
722 Factor VIII:vWF, Mab purif./Baxter
Nomenclature:
Factor VIII:vWF, Mab purif./Baxter [BIO]
Hemofil M [TR]
Antihemophilic Factor (Human), Method M, Monoclonal Purified [FDA used on insert/lableing]
Method-M [TR for immunoaffinity purification process]
Antihemophilic Factor (Human) [FDA]
Antihemophilic Factor [USAN]
Antihemophilic Factor, Human [USAN former]
Antihemophilic Factor USP [USP]
Blood-coagulation factor VIII, complex [CAS]
9001-27-8 [CAS RN]
AHF [SY]
Antihemophilic Factor VIII [SY]
Factor VIII:von Willebrand's Factor complex [SY]
FVIII:vWF complex [SY]
Haemophil, Method M [TR foreign]
FDA Class: Biologic PLA
Year of approval (FDA) = 1966
Date of 1st FDA approval = 19660311
(in format YYYYMMDD)
Index Terms:
antibodies (see also immune globulins; monoclonal antibodies)
antihemophilic factors
biopharmaceutical products
blood products
human materials used<!-- humansource -->
murine (mouse) materials used
ATCC HB 10494
Cryoprecipitated AHF (antihemophilic factor)
gentamicin (gentamycin)
murine feeder cells
Albumin (Human)
exempt from CBER lot release requirements
Factor VIII monoclonal antibodies
glycine
heat treatment (pasteurization)
histidine
immunoaffinity chromatography
lyophilized (freeze-dried)
monoclonal antibody, Factor VIII
murine monoclonal antibody, CD3
murine monoclonal antibody, Factor VIII
murine proteins
octoxynol (Triton X-100)
Plasma (Human)
polyethylene glycol (PEG)
Sepharose
tri-n-butyl phosphate (TNBP)
viral inactivation, solvent detergent
WHO Intl. Standard Factor VIII
approval dates uncertain (FDA reports erroneous, conflicting, or simply has lost the original approval dates) (FDAapproved)
EU200 Currently Approved in EU
UM001 Marketed Product in US
US200 Currently Approved in US
EM001 Marketed Product in EU
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