Essais de purification de l'insulin-like growth factor binding protein-3 (IGFBP-3) bovine

Animal productions are the results of interactions between the genetic potential of the animal, environmental factors and nutrient supplies. These interactions modulate the endocrine system, especially the somatotrope axis. IGFBP-3 is the major carrier of the IGFs (the main growth-promoting peptide) in plasma circulating as part of a 150 kDa ternary complex with one IGF peptide and a single acid-labile subunit. The IGFBP-3 purification would allow the measure of the changes in plasma IGFBP-3 throughout the day and the development and in various physiological situations. We try to purify the bovine IGFBP-3. Firstly, we load 0,5 ml of serum onto an Hiload 16/6 Superdex 200 prep grade column, then onto an ion exchange column and finally onto an IGF-I affinity column. The purification fails perhaps because the sample is to small. Secondly, 5 ml of serum is fractionated by 3 gel filtrations. Between the first and the second filtration, the fractions containing IGF-I binding activity are pooled and concentrated. Before the third gel filtration, the fractions containing this activity are pooled anew, acidified with HCl and incubated at 4 °C for 2 hours. The third gel filtration fractions are pooled and subjected to an heparin-affinity chromatography. The elution pic shows only IGFBP-3 activity. In conclusion, our purified product contains other proteins. But the ternary complex is dissociated, so an IGF-I affinity chromatography would purify the IGFBP-3 from the elution pic of the heparin-affinity column.

Grantor dc:publisher

ULiège. GxABT - Liège Université. Gembloux Agro-Bio Tech

Year dc:date

2003

Author dc:creator

• Colinet, Frédéric

Contributors dc:contributor

• Portetelle, Daniel
• Renaville, Robert

• Life sciences
• Biochemistry, biophysics & molecular biology
• Sciences du vivant
• Biochimie, biophysique & biologie moléculaire

Statement dc:rights

• restricted access
• info:eu-repo/semantics/restrictedAccess

Licence

• http://purl.org/coar/access_right/c_16ec

Language dc:language

fr

Identifier

info:hdl:2268/16404

OAI identifier oai:identifier

oai:orbi.ulg.ac.be:2268/16404