Skip to main content
Log in

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

  • Published:
Russian Journal of Bioorganic Chemistry Aims and scope Submit manuscript

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.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Subscribe and save

Springer+ Basic
EUR 32.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or Ebook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

REFERENCES

  1. Farmakopeinaya stat’ya predpriyatiya Institut bioorganicheskoi khimii im. M. M. Shemyakina i Yu.A. Ovchinnikova RAN (Pharmacopoeia Monograph of the Enterprise Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry RAS) FSP42-0452361302. Human insulin, 2002, Moscow.

  2. Haugland, R.P and Kang, H.C, Chemically Reactive Dipyrrometheneboron Difluoride Dyes, US Patent 4774339.

  3. Matz, M.V., in Methods Mol. Biol., Hicks, B.W, Ed., Totowa: Humana, 2003, vol. 221, pp. 103–116.

    Google Scholar 

  4. Sambrook, J., Fritsch, E.F., and Maniatis, T., Molecular Cloning: A Laboratory Manual, 2nd Ed., Cold Spring Harbor, NY: Cold Spring Harbor Lab., 1989.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

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.

Rights and permissions

Reprints and permissions

About this article

Cite this article

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

Download citation

  • Received:

  • Revised:

  • Issue Date:

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

Key words

Navigation