Purification and characterization of Alpha-Fetoprotein from the human hepatoblastoma HepG2 cell line in serum-free medium

Patrizia Carlini, Pasquale Ferranti, Francesca Polizio, Maria R. Ciriolo, Giuseppe Rotilio

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Alpha-fetoprotein (AFP) is a tumor-associated embryonic molecule whose precise biological function remains unclear. A complete definition of the physiological activities of this oncofetal protein has been severely limited, until now, by the lack of a purification procedure appropriate to obtain pure AFP in appreciable amount. The present report describes a purification procedure extremely rapid and simple and takes advantage of the well-known fact that AFP contains copper. We have developed a single-step purification procedure by immobilized copper-chelate affinity chromatography using the culture medium from human hepatoblastoma cell line HepG2 grown in the absence of serum. This method yields AFP at high purity and high yield. Purified AFP amino acid sequence, molecular mass, carbohydrate structure, and copper content were found to be in line with previous studies. Moreover, we found that the purified AFP has superoxide dismutase activity with efficiency similar to that of the native Cu, Zn SODs at physiological pH. This result may provide further support to the idea that AFP is a bifunctional protein, acting in cellular defence against oxidative stress both as a copper buffer and as a superoxide radical scavenger.

Original languageEnglish
Pages (from-to)869-878
Number of pages10
JournalBioMetals
Volume20
Issue number6
DOIs
Publication statusPublished - Dec 2007

Fingerprint

Hepatoblastoma
Serum-Free Culture Media
Hep G2 Cells
alpha-Fetoproteins
Purification
copper
Cells
cell lines
Copper
Cell Line
superoxide dismutase
carbohydrate structure
Proteins
Affinity chromatography
Oxidative stress
human cell lines
free radical scavengers
chelates
Molecular mass
affinity chromatography

Keywords

  • Alpha-fetoprotein
  • Copper
  • Human hepatoblastoma
  • Serum-free medium
  • Superoxide dismutase

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Purification and characterization of Alpha-Fetoprotein from the human hepatoblastoma HepG2 cell line in serum-free medium. / Carlini, Patrizia; Ferranti, Pasquale; Polizio, Francesca; Ciriolo, Maria R.; Rotilio, Giuseppe.

In: BioMetals, Vol. 20, No. 6, 12.2007, p. 869-878.

Research output: Contribution to journalArticle

Carlini, Patrizia ; Ferranti, Pasquale ; Polizio, Francesca ; Ciriolo, Maria R. ; Rotilio, Giuseppe. / Purification and characterization of Alpha-Fetoprotein from the human hepatoblastoma HepG2 cell line in serum-free medium. In: BioMetals. 2007 ; Vol. 20, No. 6. pp. 869-878.
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