Improved method for the immobilization of heparin.
The optimal conditions for immobilizing heparin through its terminal formyl group were investigated. When Amino Sepharose (1 g) was suspended in 1 ml of phosphate buffer (pH 7) containing 30 mg of heparin and 3 mg of sodium cyanoborohydride, with shaking at room temperature, the maximum immobilization of heparin (10 mg of heparin per gram of wet gel) was reached within 2 days. The Heparin Sepharose thus obtained was stable: no significant loss of the heparin content was observed after storage for 4 months at 4 degrees C. Heparin was also immobilized by the same method with Amino TSK gel G5000PW instead of Amino Sepharose 4B and was successfully applied to the high-performance liquid affinity chromatography of fibronectin and thrombin.
Sasaki
H
H
Department of Chemistry, Faculty of Science, Ochanomizu University, Tokyo, Japan.
Hayashi
A
A
Kitagaki-Ogawa
H
H
Matsumoto
I
I
Seno
N
N
D002851:Chromatography, High Pressure Liquid; D005353:Fibronectins / Q000032:analysis; D005782:Gels; D006493:Heparin / Q000302:isolation & purification*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D013917:Thrombin / Q000032:analysis
D005353:Fibronectins; D005782:Gels; D006493:Heparin; D013917:Thrombin