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Protein Expr Purif. 1999 Oct;17(1):146-52.

Plasma fibronectin: three steps to purification and stability.

Author information

1
Equipe de Recherche sur les Relations Matrice-Extracellulaire Cellules (ERRMECe), Université de Cergy-Pontoise, 2 avenue Adolphe Chauvin, Cergy-Pontoise Cedex, 95302, France.

Abstract

Large amounts of soluble fibronectin were easily purified from cryoprecipitated or fresh citrated human blood plasma by a three-step combination of gelatin and heparin-cellufine affinity chromatography. The elution conditions were optimized to obtain a homogeneous fraction on SDS-PAGE and Western blot under reducing condition. No proteolytic activities were detected by zymography at acidic or neutral pH. Furthermore, the fibronectin preparation was stable over time up to 456 h at 37 degrees C in the presence of calcium, zinc, or mercury. This preparation of very stable fibronectin, called highly purified fibronectin (hpFN), gave a yield of 7.00 +/- 0.77 mg of fibronectin per gram of cryoprecipitated plasma and 0.16 mg of fibronectin per milliliter of fresh citrated, giving a yield of 32 to 53% (from presumed fibronectin concentration). This preparation may be useful for cellular tests and interaction analysis.

PMID:
10497080
DOI:
10.1006/prep.1999.1103
[Indexed for MEDLINE]

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