Novel recombinant human lactoferrin: Differential activation of oxidative stress related gene expression

Marian L. Kruzel, Jeffrey K. Actor, Michał Zimecki, Jasen Wise, Paulina Płoszaj, Shaper Mirza, Mark Kruzel, Shen An Hwang, Xueqing Ba, Istvan Boldogh

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24 Scopus citations

Abstract

Lactoferrin, an iron-binding protein found in high concentrations in mammalian exocrine secretions, is an important component of the host defense system. It is also a major protein of the secondary granules of neutrophils from which is released upon activation. Due to its potential clinical utility, recombinant human lactoferrin (rhLF) has been produced in various eukaryotic expression systems; however, none of these are fully compatible with humans. Most of the biopharmaceuticals approved by the FDA for use in humans are produced in mammalian expression systems. The Chinese hamster ovary cells (CHO) have become the system of choice for proteins that require post-translational modifications, such as glycoproteins. The aim of this study was to scale-up expression and purification of rhLF in a CHO expression system, verify its glycan primary structure, and assess its biological properties in cell culture models. A stable CHO cell line producing >200. mg/L of rhLF was developed and established. rhLF was purified by a single-step cation-exchange chromatography procedure. The highly homogenous rhLF has a molecular weight of approximately 80. kDa. MALDI-TOF mass spectrometric analysis revealed N-linked, partially sialylated glycans at two glycosylation sites, typical for human milk LF. This novel rhLF showed a protective effect against oxidative stress in a similar manner to its natural counterpart. In addition, rhLF revealed a modulatory effect on cellular redox via upregulation of key antioxidant enzymes. These data imply that the CHO-derived rhLF is fully compatible with the native molecule, thus it has promise for human therapeutic applications.

Original languageEnglish (US)
Pages (from-to)666-675
Number of pages10
JournalJournal of Biotechnology
Volume168
Issue number4
DOIs
StatePublished - Dec 2013

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Keywords

  • Antioxidant
  • Gene expression
  • Inflammation
  • Recombinant human lactoferrin

ASJC Scopus subject areas

  • Biotechnology
  • Applied Microbiology and Biotechnology

Cite this

Kruzel, M. L., Actor, J. K., Zimecki, M., Wise, J., Płoszaj, P., Mirza, S., Kruzel, M., Hwang, S. A., Ba, X., & Boldogh, I. (2013). Novel recombinant human lactoferrin: Differential activation of oxidative stress related gene expression. Journal of Biotechnology, 168(4), 666-675. https://doi.org/10.1016/j.jbiotec.2013.09.011