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Glycoform Heterogeneity of Human Serum α1-Acid Glycoprotein Determined by CZE in Malignant Diseases

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Abstract

A novel capillary zone electrophoresis method was developed to investigate the glycoform heterogeneity of human serum α1-acid glycoprotein (AGP). The simultaneous application of a dimethyl polysiloxane coated capillary and oligoamine additives, particularly spermidine resulted in a more detailed separation of AGP glycoforms than reported previously. The relative distribution of AGP glycoforms in CZE was determined by baseline integration of peak areas and verified by peak-fitting analysis. Providing high purity of AGP samples suitable for CZE a schedule of isolation and purification steps including sample preparation and an improved technique of ion exchange chromatography was applied. Based on data obtained by CZE and on the serum AGP levels measured the serum concentrations of AGP glycoforms were calculated in cancer patients with Hodgkin and non-Hodgkin lymphoma, ovary carcinoma and melanoma compared to healthy donors. Results presented here demonstrated a significant increase in the serum concentration of the more acidic AGP fractions also indicating the overproduction of these glycoforms in cancer. In conclusion, our observations may raise the clinical diagnostic relevance of changes in the molecular heterogeneity of AGP detected by CZE in the various forms of malignant diseases.

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Acknowledgments

Authors gratefully acknowledge the technical assistance of Zsuzsa Ary and Rita Vukovics (Biochemical Department of the National Institute of Oncology). This work was supported by the grants of OTKA TO49721 and NKFP 1/A/005/04 (Medichem2 project).

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Authors and Affiliations

  1. Institute of Biomolecular Chemistry, Chemical Research Center, Hungarian Academy of Sciences, Budapest, Hungary

    Krisztina Németh, Tibor Kremmer & Júlia Visy

  2. Department of Biochemistry, National Institute of Oncology, Budapest, Hungary

    Tibor Kremmer

  3. Institute of Structural Chemistry, Chemical Research Center, Hungarian Academy of Sciences, Budapest, Hungary

    László Kocsis

Authors
  1. Krisztina Németh
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  2. Tibor Kremmer
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  3. László Kocsis
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  4. Júlia Visy
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Correspondence to Tibor Kremmer.

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Németh, K., Kremmer, T., Kocsis, L. et al. Glycoform Heterogeneity of Human Serum α1-Acid Glycoprotein Determined by CZE in Malignant Diseases. Chroma 69, 1307–1313 (2009). https://doi.org/10.1365/s10337-009-1046-7

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