Plasma fibronectin: three steps to purification and stability.

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.

journal_name

Protein Expr Purif

authors

Poulouin L,Gallet O,Rouahi M,Imhoff JM

doi

10.1006/prep.1999.1103

keywords:

subject

Has Abstract

pub_date

1999-10-01 00:00:00

pages

146-52

issue

1

eissn

1046-5928

issn

1096-0279

pii

S1046-5928(99)91103-8

journal_volume

17

pub_type

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