Status: marketed in Europe, not U.S.

Organizations involved:

Stichting Sanquin Bloedvoorziening – Manuf; R&D; Tech.; Europe mark.

New York Blood Center – Tech.

Cross ref.: See the Factor IX Products entry.

Description: Nonafact is a lyophilized (freeze-dried) formulation of highly purified Factor IX with manufacture including immunoaffinity chromatography using a matrix-bound Factor IX murine monoclonal antibody (CLB-FIX D4), solvent detergent virus inactivation, and further virus removal by nanofiltration. Nonafact is packaged in vials containing 500 IU or 1,000 IU Factor IX for reconstitution with 5 or 10 mL Water for Injection, respectively. Excipients include histidine (15 mmol/L), sucrose (151 mmol/L), and sodium chloride (75 mmol/L).

Nomenclature: Factor IX/Sanquin [BIO]; Nonafact [TR]; Coagulation Factor IX (Human) [FDA, if approved in U.S.]; 9001-28-9 [CAS RN]; Christmas factor [SY]

Companies.: Nonafact was developed and is manufactured and marketed in European countries by Stichting Sanquin Bloedvoorziening (Sanquin; the Netherlands).

Manufacture: The Plasma used for manufacture is obtained from unpaid volunteer Dutch donors at the blood centers of Stichting Sanquin Bloedvoorziening (Sanquin). The CLB subsidiary of Sanquin performs all subsequent manufacture.

The starting material is Fresh Frozen Plasma. This is subjected to cryoprecipitation. The resulting coagulation factor-depleted plasma is adsorbed onto an ion exchange chromatography matrix, washed, and Factor IX-containing “3F-concentrate” is eluted. This is diafiltered, concentrated, and stored at ≤25˚C. The 3F-concentrate is thawed and subjected to solvent-detergent viral inactivation using tri-N-butyl phosphate (TNBP) and polysorbate 80 (Tween 80) for at least 13 hours (See the Plasma SD entry for further information about solvent detergent viral inactivation). Factor IX is purified by immunoaffinity purification chromatography using murine monoclonal antibody CLB-FIX D4, a murine monoclonal antibody with specificity for intact Factor IX, and is eluted from the column using a chaotropic elution buffer. Eluate is subjected to dual nanofiltration (15 nm pore size filter), and subjected to a third chromatography step (unspecified) to remove any residual mouse protein (from the antibody used), chaotropic agents, and degraded Factor IX. Following elution for the column, Factor IX is concentrated and stored at ≤25˚C. The resulting product has a specific activity of over 200 IU/mg protein, much higher than European Pharmacopeia standards. Traces of high molecular weight proteins are present, but these do not interact with Factor IX, and no Factor IX aggregates are present. Formulated bulk is sterile filtered (0.22 µm filter), and vials are filled and lyophilized.

The CLB-FIX D4 monoclonal antibody is specific for non-activated, intact Factor IX (not cleaved at position Arg 145). CLB-FIX D4 is cultured in vitro (fermentation) using serum-free medium, followed by affinity chromatography, cation exchange chromatography, and gel filtration. The eluate is subjected to solvent-detergent viral inactivation.

Status: European Union approval (MAA) was granted on July 3, 2001.

Indications: [European Union]:

Treatment and prophylaxis of bleeding in patients with haemophilia B (congenital factor IX deficiency).

Tech. transfer: Solvent detergent viral inactivation was developed by and nonexclusively licensed from the New York Blood Center. For example, see 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).