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Large scale preparation and purification of apo-D-aminoacid oxidase for use in novel amplification assays

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Biotechnology Techniques

Summary

Dissociation of FAD from D-aminoacid oxidase occurred most rapidly at pH 6.0 in the presence of 1 M KBr. Diafiltration of 0.6 g of enzyme under these conditions yielded apoenzyme containing 1.3% of residual holoenzyme activity, which was subsequently reduced to less than 0.01% by chromatography on Blue Sepharose and ion exchange, giving material containing <1 ppb of contaminating phosphatase and nucleotidase.

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References

  • Brumby, P.E. and Massey, V. (1968)Biochem. Prep. 12, 29–41.

    CAS  Google Scholar 

  • Harbron, S., Eggelte, H.J., Fisher, M., Benson, M. and Rabin, B.R. (1992) submitted for publication inClin. Chem.

  • Leonil, J., Langrene, S., Sicsic, S. and Le Goffic, F. (1985).J. Chromatography 347, 316–319.

    Article  CAS  Google Scholar 

  • Lindner, N.N., Jeffcoat, R. and Lowe, C.R. (1989)J. Chromatography 473, 227–240.

    Article  CAS  Google Scholar 

  • Massey, V. and Curti, B. (1966)J. Biol. Chem. 241, 3417–3423.

    CAS  Google Scholar 

  • Obzansky, D.M., Rabin, B.R., Simons, D.M., Tseng, S.Y., Severino, D.M., Eggelte, H.J., Fisher, M., Harbron, S., Stout, R.W., and Di Paolo, M.J. (1991)Clin. Chem. 37, 1513–1518.

    CAS  Google Scholar 

  • Rabin, B.R., Harbron, S. and Eggelte, H.J. (1990) World Patent Application Number 90/01559.

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Authors and Affiliations

  1. London Biotechnology Ltd, Department of Biochemistry and Molecular Biology, University College London, Gower Street, WC1E 6BT, London, UK

    Stuart Harbron, Mark Fisher & Brian R. Rabin

Authors
  1. Stuart Harbron
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  2. Mark Fisher
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  3. Brian R. Rabin
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Harbron, S., Fisher, M. & Rabin, B.R. Large scale preparation and purification of apo-D-aminoacid oxidase for use in novel amplification assays. Biotechnol Tech 6, 55–60 (1992). https://doi.org/10.1007/BF02438689

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  • DOI: https://doi.org/10.1007/BF02438689

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