High Level Expression of Active Human Prothrombin in a Vaccina Virus Expression System

F G Falkner; P L Turecek; R T A MacGillivray; W Bodemer; F Scheiflinger; S Kandels; A Mitterer; O Kistner; N Barrett; J Eibl; F Dorner

doi:10.1055/s-0038-1656335

Thrombosis and Haemostasis, Inhaltsverzeichnis

Thromb Haemost 1992; 68(02): 119-124
DOI: 10.1055/s-0038-1656335

Original Article

Schattauer GmbH Stuttgart

High Level Expression of Active Human Prothrombin in a Vaccina Virus Expression System

F G Falkner

The Immuno AG, Biomedical Research Center, Orth/Donau, Austria, and the University of British Columbia, Department of Biochemistry, Vancouver, B. C., Canada

,

P L Turecek

The Immuno AG, Biomedical Research Center, Orth/Donau, Austria, and the University of British Columbia, Department of Biochemistry, Vancouver, B. C., Canada

,

R T A MacGillivray

The Immuno AG, Biomedical Research Center, Orth/Donau, Austria, and the University of British Columbia, Department of Biochemistry, Vancouver, B. C., Canada

,

W Bodemer

The Immuno AG, Biomedical Research Center, Orth/Donau, Austria, and the University of British Columbia, Department of Biochemistry, Vancouver, B. C., Canada

,

F Scheiflinger

The Immuno AG, Biomedical Research Center, Orth/Donau, Austria, and the University of British Columbia, Department of Biochemistry, Vancouver, B. C., Canada

,

S Kandels

The Immuno AG, Biomedical Research Center, Orth/Donau, Austria, and the University of British Columbia, Department of Biochemistry, Vancouver, B. C., Canada

,

A Mitterer

The Immuno AG, Biomedical Research Center, Orth/Donau, Austria, and the University of British Columbia, Department of Biochemistry, Vancouver, B. C., Canada

,

O Kistner

The Immuno AG, Biomedical Research Center, Orth/Donau, Austria, and the University of British Columbia, Department of Biochemistry, Vancouver, B. C., Canada

,

N Barrett

The Immuno AG, Biomedical Research Center, Orth/Donau, Austria, and the University of British Columbia, Department of Biochemistry, Vancouver, B. C., Canada

,

J Eibl

The Immuno AG, Biomedical Research Center, Orth/Donau, Austria, and the University of British Columbia, Department of Biochemistry, Vancouver, B. C., Canada

,

F Dorner

The Immuno AG, Biomedical Research Center, Orth/Donau, Austria, and the University of British Columbia, Department of Biochemistry, Vancouver, B. C., Canada

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Abstract

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Summary

We have worked out an efficient and time saving procedure for the expression of recombinant human prothrombin. The glycoprotein was expressed in the vaccinia virus expression system in several mammalian cell lines. The kidney cell lines Vero and BHK and the human cell line Hela were found to efficiently secrete prothrombin. Expression levels of 3–4 µg of factor II per 10⁶ cells per day corresponding to 18–23 mU per 10⁶ cells per day were achieved. Since the expression levels obtained with the vaccinia virus/Vero cell system were comparable to those obtained in amplified transformed CHO cells it provides an alternative system for the efficient expression of human prothrombin and may allow to further elucidate structure-function relationships of (pro)thrombin and its various effectors.

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Referenzen

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