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Biotechnology Techniques

, Volume 11, Issue 9, pp 697–700 | Cite as

Microquantification of proteins with low chromophore content

  • W. Quintilio
  • R.A. Sato
  • A.M. da Silveira
  • G. Barratt
  • E. Fattal
  • M.H. Bueno-da-Costa
Article

Abstract

The advantages of the intrinsic fluorescence of the tryptophan and the absorbance of the methionine residues of the 18 kDa-hsp - a recombinant protein from Mycobacterium leprae - was exploited here to develop a sensitive and low costs method for protein assaying. They presented linearity between 3 and 1000 μg of protein. The correlations between intrinsic fluorescence or absorbance at 230 nm and protein contents were both superiors to 0.99. These methods can be extended to others proteins with low aromatic residue contents.

Keywords

Protein Content Tryptophan Methionine Recombinant Protein Mycobacterium 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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References

  1. Anicetti, V and Hancock, W (1993). In: « Protein Purification Process Engineering », RG Harrison, ed. pp 11–35, Marcel Dekker Inc., New York, USAGoogle Scholar
  2. Booth, RJ, Grandson, PM, Prestidge, RL and Watson, JD (1988). Immunol. Letters 19: 65–70Google Scholar
  3. Costa, MHB, Ueda, CMPM, Sato, RA, Liberman, C and Raw, I (1995). Biotech. Techniques 9: 527–532Google Scholar
  4. Dwyer, J.H. (1993). Process purification. In Protein Biotechnology. Isolation, Characterisation, and Stabilisation, Felix Franks, ed. pp 533–572, Humana Press, Totowa, New Jersey, USAGoogle Scholar
  5. Lamb, FI, Singh, NB and Colston, MJ (1992). J. Immunol. 144: 1922–1925Google Scholar
  6. Peterson, GL (1983). Meth. Enzymol. 91: 95–116Google Scholar
  7. Sato, RA and Costa, MHC (1996). Biotech. Letters 18: 275–280Google Scholar
  8. Stoscheck, C (1990). Guide to protein purification, In: Methods in Enzymology, Murray P. Deutscher, ed, vol 182, pp 50–68, New York, USA, Academic Press, Inc.Google Scholar

Copyright information

© Chapman and Hall 1997

Authors and Affiliations

  • W. Quintilio
    • 1
  • R.A. Sato
    • 1
  • A.M. da Silveira
    • 2
  • G. Barratt
    • 2
  • E. Fattal
    • 2
  • M.H. Bueno-da-Costa
    • 1
    • 2
  1. 1.Lab. Microesferas e Lipossomos, C. de BiotecnologiaButantan, Sao Paulo., S.P.Brasil
  2. 2.Centre D‘Etudes PharmaceutiquesUniversité Paris XI, URA-CNRS 1218Chatenay-Malabry CedexFrance

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