Rapid methods for isolation of human plasma fibronectin.
Simplified procedures have been developed for isolation of human plasma fibronectin by affinity chromatography on gelatin-agarose. In one method, fibronectin is eluted with 3 M urea, and this reagent is quickly removed by adsorbing the protein onto heparin-agarose, followed by 0.4 M NaCl elution. In a shorter process, fibronectin is eluted from gelatin-agarose simply by decreasing the buffer pH below 6. After lyophilization the purified protein can be readily dissolved in water. The fraction not adsorbed to gelatin can be used to purify other proteins, including factor VIII whose procoagulant activity is quantitatively recovered.
D011232:Chemical Precipitation; D002846:Chromatography, Affinity / Q000379:methods; D003450:Cryoglobulins / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D005169:Factor VIII / Q000032:analysis; D005353:Fibronectins / Q000097:blood / Q000302:isolation & purification*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D011487:Protein Conformation; D014508:Urea
D003450:Cryoglobulins; D005353:Fibronectins; D014508:Urea; D005169:Factor VIII