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

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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.

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Authors and Affiliations

  1. Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya 16/10, 117997, Moscow, Russia

    A. N. Aleksandrov, Yu. S. Skoblov, E. D. Shibanova, D. I. Bairamashvili & A. I. Miroshnikov

  2. Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, ul. Vavilova 32, 119991, Moscow, Russia

    M. Yu. Skoblov

Authors
  1. A. N. Aleksandrov
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  2. Yu. S. Skoblov
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  3. M. Yu. Skoblov
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  4. E. D. Shibanova
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  5. D. I. Bairamashvili
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  6. A. I. Miroshnikov
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Corresponding author

Correspondence to A. N. Aleksandrov.

Additional information

Translated from Bioorganicheskaya Khimiya, Vol. 31, No. 1, 2005, pp. 73–76.

Original Russian Text Copyright © 2005 by Aleksandrov, Yu. Skoblov, M. Skoblov, Shibanova, Bairamashvili, Miroshnikov.

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Aleksandrov, A.N., Skoblov, Y.S., Skoblov, M.Y. et al. A PCR-based semiquantitative assay of DNA impurities in recombinant protein preparations. Russ J Bioorg Chem 31, 66–69 (2005). https://doi.org/10.1007/s11171-005-0008-6

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  • DOI: https://doi.org/10.1007/s11171-005-0008-6

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