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Simplified purification of equine chorionic gonadotropin (eCG)––an example of the use of magnetic microsorbents for the isolation of glycoproteins from serum

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Abstract

Classical purification of the glycoprotein equine chorionic gonadotropin (eCG) from serum includes pH fractionation with metaphosphoric acid, two ethanol precipitation steps as well as dialysis followed by fixed-bed chromatography. A simplified process requiring only 1/3 of the solvent and improving the yield from 53 to 65% has been developed. The process comprises an ultra-/diafiltration step after the first ethanol precipitation, directly followed by an adsorption/desorption procedure based on magnetic microadsorbents with N,N-diethyl-ammonium functionalization. The process reaches an overall purification factor of eCG of more than 1800 and an average product activity of 1300 IUELISA/mg. After adapting the parameters of the fractionation and the type of magnetic microadsorbents, the new concept is likely to be transferable to other serum proteins.

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Notes

  1. A fixed correlation exists between the biological activities determined in mice or rats: 3.8 mu/ml = 1 IU/ml. Therefore the used serum corresponded to an average of 53 IU/ml.

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Acknowledgments

The authors would like to thank their industrial partners (ABBIS, INLAB GmbH) of the joint project for the fruitful discussion. We also thank chemagen Biopolymer-Technologie AG in Baesweiler for the preparation of the carboxylated magnetic polyvinyl alcohol beads. Funding by the Federal Ministry of Economics and Technology (BMWi) is acknowledged.

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Authors and Affiliations

  1. Institute of Functional Interfaces, Karlsruhe Institute of Technology, Hermann von Helmholtz Platz 1, 76344, Eggenstein Leopoldshafen, Germany

    Christine Müller & Matthias Franzreb

  2. Biomaterials Department, INNOVENT e. V., Prüssingstrasse 27b, 07745, Jena, Germany

    Kerstin Wagner

  3. fzmb GmbH Institute for Medical Technology and Biotechnology, Geranienweg 7, 99947, Bad Langensalza, Germany

    Katrin Frankenfeld

Authors
  1. Christine Müller
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  2. Kerstin Wagner
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  3. Katrin Frankenfeld
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  4. Matthias Franzreb
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Correspondence to Matthias Franzreb.

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Müller, C., Wagner, K., Frankenfeld, K. et al. Simplified purification of equine chorionic gonadotropin (eCG)––an example of the use of magnetic microsorbents for the isolation of glycoproteins from serum. Biotechnol Lett 33, 929–936 (2011). https://doi.org/10.1007/s10529-010-0512-5

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  • DOI: https://doi.org/10.1007/s10529-010-0512-5

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