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Analysis of expression kinetics and activity of a new B-domain truncated and full-length FVIII protein in three different cell lines

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Abstract.

Transient expression of full-length wild-type (wt) and a new B-domain truncated (ΔB) FVIII has been investigated in three eukaryotic cell lines (HEK-293, COS, CHO). When expressed in CHO cells, both FVIII proteins reached the same peak antigen levels, whereas in HEK-293 and COS cells those of FVIII/ΔB were up to sixfold those of FVIII/wt. Investigation of specific activity of the recombinant FVIII proteins using a chromogenic and a one-stage assay in addition to the FVIII-antigen ELISA revealed large variations: In HEK-293 cells specific activity of FVIII/ΔB measured with both assays was higher than that of FVIII/wt. In COS cells specific activity of both FVIII proteins was higher measured in the one-stage assay than in the chromogenic assay. In CHO cells both FVIII proteins had similar specific activity in each assay. In summary, expression kinetics and specific activity of conditioned medium vary depending on cell type used.

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Authors and Affiliations

  1. Institute for Experimental Hematology and Transfusion Medicine, University of Bonn, Sigmund-Freud-Strasse 25, D-53105 Bonn, Germany Tel.: 0049-228-287-6751, Fax: 0049-228-287-4320, , , , , , DE

    A. Haack, C. Schmitt, P. Hanfland, H.-H. Brackmann & R. Schwaab

  2. Department of Cardiology and Pneumology, University Hospital Benjamin Franklin, Free University of Berlin, Hindenburgdamm 30, D-12200 Berlin, Germany, , , , , , DE

    W. Poller

  3. Institute of Human Genetics, University of Würzburg, Am Hubland, D-97074 Würzburg, Germany, , , , , , DE

    J. Oldenburg

Authors
  1. A. Haack
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  2. C. Schmitt
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  3. W. Poller
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  4. J. Oldenburg
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  5. P. Hanfland
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  6. H.-H. Brackmann
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  7. R. Schwaab
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Additional information

Received: June 4, 1998 / Accepted: December 15, 1998

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Haack, A., Schmitt, C., Poller, W. et al. Analysis of expression kinetics and activity of a new B-domain truncated and full-length FVIII protein in three different cell lines. Ann Hematol 78, 111–116 (1999). https://doi.org/10.1007/s002770050486

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  • DOI: https://doi.org/10.1007/s002770050486

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