Performing a Separation of Coagulation Factors with GE Healthcare Products Based on Heparin

Extracted from Affinity Chromatography Principles and Methods, GE Healthcare, 2007

Blood coagulation factors form an extremely important group of proteins for research, medical and clinical applications. The information about the purification of DNA binding proteins (page 60, DNA binding proteins) is applicable also to the purification of coagulation factors.

Purification Examples

Purification of antithrombin III from bovine plasma on HiTrap Heparin HP, 1 ml

Fig. 38. Purification of antithrombin III from bovine plasma on HiTrap Heparin HP, 1 ml.

 

Purification of antithrombin-III from human plasma on Heparin Sepharose 6 Fast Flow

Fig. 39. Purification of antithrombin-III from human plasma on Heparin Sepharose 6 Fast Flow. Peak B elutes with wash buffer. Peak C elutes with elution buffer and includes a more active form of antithrombin-III.

Performing a Separation

As for DNA binding proteins, see page 63 (Performing a Separation of DNA binding proteins with GE Healthcare Products Based on Heparin).

Since the heparin acts as an affinity ligand for coagulation factors, it may be advisable to include a minimum concentration of 0.15 M NaCl in the binding buffer.

If an increasing salt gradient gives unsatisfactory results, use heparin (1–5 mg/ml) as a competing agent in the elution buffer.

Materials

     

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