A highly purified factor VIII: c concentrate prepared from cryoprecipitate by ion-exchange chromatography

T. Burnouf, M. Burnouf-Radosevich, J. J. Huart, M. Goudemand

Research output: Contribution to journalArticle

53 Citations (Scopus)

Abstract

A new ion-exchange chromatographic procedure has been developed to produce a highly purified factor VIII (FVIII) concentrate from plasma cryoprecipitate. Solubilized cryoprecipitate, after adsorption on aluminium hydroxide and cold precipitation, was treated with 0.3% tri(n-butyl)phosphate and 1% Tween 80 at 25°C for at least 8 h to inactivate lipid-enveloped viruses. The fraction was then loaded onto a column packed with DEAE-Fractogel TSK 650 M and chromatographed. Most proteins and TnBP-Tween 80 flowed through the gel unretarded. FVIII:c, which bound to the gel, was eluted by increasing the ionic strength, then was directly filter-sterilized without ultrafiltration or addition of a protein stabilizer. Chromatographic recovery of FVIII:c was 80-90%. After freeze-drying, FVIII:c was at a concentration of 42.5 ± 9.5 IU/ml and had a specific activity of 175.4 ± 37.8 IU/mg (n = 40), corresponding to a purification factor of over 12,000 from plasma. The typical yield of the freeze-dried FVIII:c from cryoprecipitate was 55-65%. FVIII:c was stable for over 24 h at room temperature in the liquid state. The mean content of fibrinogen and immunoglobulin G was only 65 and 100 mg/ml, respectively, corresponding to 1.4 and 2.3 mg/1,000 IU FVIII:c. This concentrate, which is much purer than traditional FVIII concentrates, has been found to be well tolerated and effective in clinical treatment of hemophilia A patients.

Original languageEnglish
Pages (from-to)8-15
Number of pages8
JournalVox Sanguinis
Volume60
Issue number1
Publication statusPublished - 1991
Externally publishedYes

Fingerprint

Ion Exchange Chromatography
Factor VIII
Polysorbates
Gels
Aluminum Hydroxide
Freeze Drying
Ion Exchange
Ultrafiltration
Hemophilia A
Osmolar Concentration
Fibrinogen
Adsorption
Proteins
Immunoglobulin G
Viruses
Lipids
Temperature

ASJC Scopus subject areas

  • Hematology

Cite this

A highly purified factor VIII : c concentrate prepared from cryoprecipitate by ion-exchange chromatography. / Burnouf, T.; Burnouf-Radosevich, M.; Huart, J. J.; Goudemand, M.

In: Vox Sanguinis, Vol. 60, No. 1, 1991, p. 8-15.

Research output: Contribution to journalArticle

Burnouf, T, Burnouf-Radosevich, M, Huart, JJ & Goudemand, M 1991, 'A highly purified factor VIII: c concentrate prepared from cryoprecipitate by ion-exchange chromatography', Vox Sanguinis, vol. 60, no. 1, pp. 8-15.
Burnouf, T. ; Burnouf-Radosevich, M. ; Huart, J. J. ; Goudemand, M. / A highly purified factor VIII : c concentrate prepared from cryoprecipitate by ion-exchange chromatography. In: Vox Sanguinis. 1991 ; Vol. 60, No. 1. pp. 8-15.
@article{ae35f1f6950f4d4abbdc970546410220,
title = "A highly purified factor VIII: c concentrate prepared from cryoprecipitate by ion-exchange chromatography",
abstract = "A new ion-exchange chromatographic procedure has been developed to produce a highly purified factor VIII (FVIII) concentrate from plasma cryoprecipitate. Solubilized cryoprecipitate, after adsorption on aluminium hydroxide and cold precipitation, was treated with 0.3{\%} tri(n-butyl)phosphate and 1{\%} Tween 80 at 25°C for at least 8 h to inactivate lipid-enveloped viruses. The fraction was then loaded onto a column packed with DEAE-Fractogel TSK 650 M and chromatographed. Most proteins and TnBP-Tween 80 flowed through the gel unretarded. FVIII:c, which bound to the gel, was eluted by increasing the ionic strength, then was directly filter-sterilized without ultrafiltration or addition of a protein stabilizer. Chromatographic recovery of FVIII:c was 80-90{\%}. After freeze-drying, FVIII:c was at a concentration of 42.5 ± 9.5 IU/ml and had a specific activity of 175.4 ± 37.8 IU/mg (n = 40), corresponding to a purification factor of over 12,000 from plasma. The typical yield of the freeze-dried FVIII:c from cryoprecipitate was 55-65{\%}. FVIII:c was stable for over 24 h at room temperature in the liquid state. The mean content of fibrinogen and immunoglobulin G was only 65 and 100 mg/ml, respectively, corresponding to 1.4 and 2.3 mg/1,000 IU FVIII:c. This concentrate, which is much purer than traditional FVIII concentrates, has been found to be well tolerated and effective in clinical treatment of hemophilia A patients.",
author = "T. Burnouf and M. Burnouf-Radosevich and Huart, {J. J.} and M. Goudemand",
year = "1991",
language = "English",
volume = "60",
pages = "8--15",
journal = "Vox Sanguinis",
issn = "0042-9007",
publisher = "Blackwell Publishing Ltd",
number = "1",

}

TY - JOUR

T1 - A highly purified factor VIII

T2 - c concentrate prepared from cryoprecipitate by ion-exchange chromatography

AU - Burnouf, T.

AU - Burnouf-Radosevich, M.

AU - Huart, J. J.

AU - Goudemand, M.

PY - 1991

Y1 - 1991

N2 - A new ion-exchange chromatographic procedure has been developed to produce a highly purified factor VIII (FVIII) concentrate from plasma cryoprecipitate. Solubilized cryoprecipitate, after adsorption on aluminium hydroxide and cold precipitation, was treated with 0.3% tri(n-butyl)phosphate and 1% Tween 80 at 25°C for at least 8 h to inactivate lipid-enveloped viruses. The fraction was then loaded onto a column packed with DEAE-Fractogel TSK 650 M and chromatographed. Most proteins and TnBP-Tween 80 flowed through the gel unretarded. FVIII:c, which bound to the gel, was eluted by increasing the ionic strength, then was directly filter-sterilized without ultrafiltration or addition of a protein stabilizer. Chromatographic recovery of FVIII:c was 80-90%. After freeze-drying, FVIII:c was at a concentration of 42.5 ± 9.5 IU/ml and had a specific activity of 175.4 ± 37.8 IU/mg (n = 40), corresponding to a purification factor of over 12,000 from plasma. The typical yield of the freeze-dried FVIII:c from cryoprecipitate was 55-65%. FVIII:c was stable for over 24 h at room temperature in the liquid state. The mean content of fibrinogen and immunoglobulin G was only 65 and 100 mg/ml, respectively, corresponding to 1.4 and 2.3 mg/1,000 IU FVIII:c. This concentrate, which is much purer than traditional FVIII concentrates, has been found to be well tolerated and effective in clinical treatment of hemophilia A patients.

AB - A new ion-exchange chromatographic procedure has been developed to produce a highly purified factor VIII (FVIII) concentrate from plasma cryoprecipitate. Solubilized cryoprecipitate, after adsorption on aluminium hydroxide and cold precipitation, was treated with 0.3% tri(n-butyl)phosphate and 1% Tween 80 at 25°C for at least 8 h to inactivate lipid-enveloped viruses. The fraction was then loaded onto a column packed with DEAE-Fractogel TSK 650 M and chromatographed. Most proteins and TnBP-Tween 80 flowed through the gel unretarded. FVIII:c, which bound to the gel, was eluted by increasing the ionic strength, then was directly filter-sterilized without ultrafiltration or addition of a protein stabilizer. Chromatographic recovery of FVIII:c was 80-90%. After freeze-drying, FVIII:c was at a concentration of 42.5 ± 9.5 IU/ml and had a specific activity of 175.4 ± 37.8 IU/mg (n = 40), corresponding to a purification factor of over 12,000 from plasma. The typical yield of the freeze-dried FVIII:c from cryoprecipitate was 55-65%. FVIII:c was stable for over 24 h at room temperature in the liquid state. The mean content of fibrinogen and immunoglobulin G was only 65 and 100 mg/ml, respectively, corresponding to 1.4 and 2.3 mg/1,000 IU FVIII:c. This concentrate, which is much purer than traditional FVIII concentrates, has been found to be well tolerated and effective in clinical treatment of hemophilia A patients.

UR - http://www.scopus.com/inward/record.url?scp=0026086692&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0026086692&partnerID=8YFLogxK

M3 - Article

C2 - 1905084

AN - SCOPUS:0026086692

VL - 60

SP - 8

EP - 15

JO - Vox Sanguinis

JF - Vox Sanguinis

SN - 0042-9007

IS - 1

ER -