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Russian Journal of Bioorganic Chemistry

, Volume 31, Issue 1, pp 66–69 | Cite as

A PCR-based semiquantitative assay of DNA impurities in recombinant protein preparations

  • A. N. Aleksandrov
  • Yu. S. Skoblov
  • M. Yu. Skoblov
  • E. D. Shibanova
  • D. I. Bairamashvili
  • A. I. Miroshnikov
Article

Abstract

A semiquantitative assay of DNA impurities in preparations of human recombinant insulin is described. The assay is based on the detection of a fragment of the ampicillin-resistant gene within the producer strain DNA by PCR. The analysis of PCR products of the studied preparations and PCR products containing known amounts of E. coli total DNA enabled a quantitative determination of the producer strain DNA content in the preparations under study. The sensitivity of the method is 7 pg of E. coli DNA per 10µg of human recombinant insulin. The high sensitivity of the method allows us to recommend it for the quantitative determination of DNA content in recombinant preparations that do not inhibit PCR.

Key words

DNA determination human recombinant insulin PCR 

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Copyright information

© MAIK “Nauka/Interperiodica” 2005

Authors and Affiliations

  • A. N. Aleksandrov
    • 1
  • Yu. S. Skoblov
    • 1
  • M. Yu. Skoblov
    • 2
  • E. D. Shibanova
    • 1
  • D. I. Bairamashvili
    • 1
  • A. I. Miroshnikov
    • 1
  1. 1.Shemyakin-Ovchinnikov Institute of Bioorganic ChemistryRussian Academy of SciencesMoscowRussia
  2. 2.Engelhardt Institute of Molecular BiologyRussian Academy of SciencesMoscowRussia

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