African Journal of
Biochemistry Research

  • Abbreviation: Afr. J. Biochem. Res.
  • Language: English
  • ISSN: 1996-0778
  • DOI: 10.5897/AJBR
  • Start Year: 2007
  • Published Articles: 425

Full Length Research Paper

Enrichment methods for revealing lower molecular weight biomarkers in human breast plasma proteome

Bechr Hamrita1*, Hela ben Nasr1#, Sami Remadi2, Anouar Chaïeb3 and Karim Chahed1,4   #Bechr Hamrita and Hela ben Nasr contributed equally to this study and are co-first authors.
1Laboratory of immunooncology, Faculty of Medecine, Monastir, Tunisia 2Laboratory of Cytopathology, Sousse, Tunisia. 3Department of Obstetrics, CHU Farhat Hached, Sousse, Tunisia. 4Faculty of Sciences, Sfax Tunisia.
Email: [email protected]

  •  Published: 31 January 2011


Breast cancer is the most common malignancy in women, comprising about 18% of all female cancers. The most important tools in screening and early detection are imaging techniques: mammography, ultrasonography and magnetic resonance imaging. However, up to 20% of new breast cancer incidents cannot be detected by these methods. Up to now, poor diagnosis of breast cancer is due to a lack of specific biomarkers. Therefore, there is much interest in identifying useful pathological markers that can help in detection and treatment of this disease. Up to now, detecting minor proteins in human plasma is a difficult strategy due to the presence of several highly abundant proteins such as albumin and haptoglobins. The objective of this study was to develop a reproducible method that depletes high-abundant proteins and improves detection of minor ones. For this purpose, we evaluated the impact of using Affi-gel blue affinity columns and organic solvent (acetonitrile) precipitation on the removal of high abundant proteins. Fractionated plasma protein extracts were subsequently analyzed by SDS-PAGE and two dimensional gel electrophoresis. Our findings confirm that Affi-gel blue can enhance detection of minor plasma proteins and could thus be a useful strategy in the discovery of new breast cancer markers.


Key words: Affigel blue, albumin, acetonitrile, plasma, proteomic separation, breast cancer.


2DE, Two-dimensional dlectrophoresis; SDS-PAGE, sodium dodecylsulfate-polyacrylamide gel electrophoresis; IEF, isoelectric focusing; ACN, acetonitrile; pI, isoelectric point; MW, molecular weight.