Get access

Isolation and characterization of a subset of erythropoietin glycoforms with cytoprotective but minimal erythropoietic activity

Authors

  • Mónica Mattio,

    1. Cell Culture Laboratory, School of Biochemistry and Biological Sciences, Universidad Nacional del Litoral, Ciudad Universitaria, C.C. 242 (S3000ZAA), Santa Fe, Argentina
    Search for more papers by this author
  • Natalia Ceaglio,

    1. Cell Culture Laboratory, School of Biochemistry and Biological Sciences, Universidad Nacional del Litoral, Ciudad Universitaria, C.C. 242 (S3000ZAA), Santa Fe, Argentina
    Search for more papers by this author
  • Marcos Oggero,

    Corresponding author
    1. Cell Culture Laboratory, School of Biochemistry and Biological Sciences, Universidad Nacional del Litoral, Ciudad Universitaria, C.C. 242 (S3000ZAA), Santa Fe, Argentina
    Current affiliation:
    1. Laboratorio de Cultivos Celulares, Facultad de Bioquímica y Ciencias Biológicas, Universidad Nacional del Litoral, Ciudad Universitaria, C.C. 242 (S3000ZAA), Santa Fe, Argentina
    • Laboratorio de Cultivos Celulares, Facultad de Bioquímica y Ciencias Biológicas, Universidad Nacional del Litoral, Ciudad Universitaria, C.C. 242 (S3000ZAA), Santa Fe, Argentina
    Search for more papers by this author
  • Norma Perotti,

    1. Cell Culture Laboratory, School of Biochemistry and Biological Sciences, Universidad Nacional del Litoral, Ciudad Universitaria, C.C. 242 (S3000ZAA), Santa Fe, Argentina
    Search for more papers by this author
  • Ignacio Amadeo,

    1. Cell Culture Laboratory, School of Biochemistry and Biological Sciences, Universidad Nacional del Litoral, Ciudad Universitaria, C.C. 242 (S3000ZAA), Santa Fe, Argentina
    2. Zelltek SA, RN 168 – PTLC – Paraje “El Pozo” (3000), Santa Fe, Argentina
    Search for more papers by this author
  • Gustavo Orozco,

    1. Zelltek SA, RN 168 – PTLC – Paraje “El Pozo” (3000), Santa Fe, Argentina
    Search for more papers by this author
  • Guillermina Forno,

    1. Cell Culture Laboratory, School of Biochemistry and Biological Sciences, Universidad Nacional del Litoral, Ciudad Universitaria, C.C. 242 (S3000ZAA), Santa Fe, Argentina
    2. Zelltek SA, RN 168 – PTLC – Paraje “El Pozo” (3000), Santa Fe, Argentina
    Search for more papers by this author
  • Ricardo Kratje,

    1. Cell Culture Laboratory, School of Biochemistry and Biological Sciences, Universidad Nacional del Litoral, Ciudad Universitaria, C.C. 242 (S3000ZAA), Santa Fe, Argentina
    Search for more papers by this author
  • Marina Etcheverrigaray

    1. Cell Culture Laboratory, School of Biochemistry and Biological Sciences, Universidad Nacional del Litoral, Ciudad Universitaria, C.C. 242 (S3000ZAA), Santa Fe, Argentina
    Search for more papers by this author

Abstract

Although historically used for the treatment of anemia, erythropoietin (EPO) has emerged as a neurotrophic and neuroprotective agent in different conditions of neuronal damage (traumatic brain injury, ischemia, spinal cord compression, peripheral neuropathy, retinal damage, epilepsy, Parkinson's Disease, among others). Nonetheless, EPO's therapeutic application is limited due to its hematological side-effects. With the aim of obtaining EPO derivatives resembling the hormone isolated from cells and tissues of neural origin, a novel combination of less acidic EPO glycoforms -designated as neuroepoetin (rhNEPO)- was purified to homogeneity from the supernatant of a CHO-producing cell line by a four-step chromatographic procedure. This simple and single process allowed us to prepare two EPO derivatives with distinct therapeutic expectations: the hematopoietic version and a minimally hematopoietic, but mainly in vitro cytoprotective, alternative. Further biological characterization showed that the in vivo erythropoietic activity of rhNEPO was 25-times lower than that of rhEPO. Interestingly, using different in vitro cytoprotective assays we found that this molecule exerts cytoprotection equivalent to, or better than, that of rhEPO in cells of neural phenotype. Furthermore, despite its shorter plasma half-life, rhNEPO was rapidly absorbed and promptly detected in the cerebrospinal fluid after intravenous administration in rats (5 min postinjection, in comparison with 30 min for rhEPO). Therefore, our results support the study of neuroepoetin as a potential drug for the treatment of neurological diseases, combining high cytoprotective activity with reduced hematological side-effects. © 2011 American Institute of Chemical Engineers Biotechnol. Prog., 2011

Get access to the full text of this article

Ancillary