Large-scale production and properties of a solvent-detergent-treated factor IX concentrate from human plasma

C. Michalski, F. Bal, T. Burnouf, M. Goudemand

Research output: Contribution to journalArticle

35 Citations (Scopus)

Abstract

A human solvent-detergent (SD)-treated factor IX concentrate has been produced from cryoprecipitate-poor plasma using DEAE-Sepharose CL-6B and heparin-Sepharose CL-6B chromatography. The DEAE eluate was incubated with an SD mixture [0.3% tri(n-butyl)phosphate-1% Tween 80, 6-h at 24°C] which was found to inactivate, in less than 1 h, more than 3.8 log10 of vesicular stomatitis virus and more than 4.8 log10 of Sindbis virus; the SD was removed by a subsequent heparin adsorption step. The specific activity of the concentrate was 10.9 ± 1.3 IU factor IX:c/mg protein (n=15). The factor IX coagulant to antigen ratio was 0.7 ± 0.1. The concentrate was essentially free of factors II, VII and X, and protein C. The usual major contaminants of prothrombin complex concentrate (PCC) were absent: the concentrate contained about 94% alpha-1 proteins, and only 4 major proteins were resolved by SDS-PAGE (respective apparent molecular weight: 130, 86, 76 and 69 kilodaltons) and by crossed immunoelectrophoresis against an anti-PCC serum. The nonactivated partial thromboplastin time was equivalent to that of PCC; the product was devoid of factor IXa, of other activated procoagulant factors and of coagulant-active phospholipids (removed with SD in the heparin breakthrough fraction). Animal studies using the Wessler test and acute-toxicity test in rabbits revealed no adverse side effects. SD treatment could thus be used to inactivate viruses in factor IX concentrate and improve the safety of replacement therapy in hemophilia B.

Original languageEnglish
Pages (from-to)202-210
Number of pages9
JournalVox Sanguinis
Volume55
Issue number4
Publication statusPublished - 1988
Externally publishedYes

Fingerprint

Detergents
Coagulants
Factor IX
Heparin
Factor IXa
Acute Toxicity Tests
Sindbis Virus
Two-Dimensional Immunoelectrophoresis
Viruses
Hemophilia B
Factor X
Vesicular Stomatitis
Factor VII
Proteins
Partial Thromboplastin Time
Polysorbates
Prothrombin
Protein C
Adsorption
Chromatography

ASJC Scopus subject areas

  • Hematology

Cite this

Large-scale production and properties of a solvent-detergent-treated factor IX concentrate from human plasma. / Michalski, C.; Bal, F.; Burnouf, T.; Goudemand, M.

In: Vox Sanguinis, Vol. 55, No. 4, 1988, p. 202-210.

Research output: Contribution to journalArticle

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