heparin sepharose 6FF 说明书.pdf

Heparin Sepharose 6 Fast Flow Instructions 71-7190-00 AE Affinity media GE Healthcare Heparin is a naturally occurring glycosaminoglycan which serves as an effective affinity binding and ion exchange ligand for a wide range of biomolecules, including coagulation factors and other plasma proteins, lipoproteins, protein synthesis factors, enzymes that act on nucleic acids and steroid receptors. By coupling heparin to SepharoseTM 6 Fast Flow with a chemically optimized linkage, GE Healthcare provides an excellent medium for both laboratory and process scale affinity purification. p. 2 Contents 1. Medium characteristics 3 2. Packing columns 4 3. Evaluation of packing 14 4. Operation 17 5. Maintenance 18 6. Ordering information 20 1. Medium characteristics Heparin is a glycosaminoglycan consisting of alternating hexuronic (D-glucuronic or L-iduronic) and D-glucosamine residues. The polymer is heavily sulphated, carrying sulphamino (N-sulphate) groups at C-2 of the glucosamine units as well as ester sulphate (O-sulphate) groups in various positions. (Figure 1). The heparin ligand used in Heparin Sepharose 6 Fast Flow is isolated from porcine intestinal mucosa, and has a mole cular weight distribution over the range 5 000–30 000. The base matrix, Sepharose 6 Fast Flow, consists of highly cross-linked 6% agarose beads. It provides the medium with high physical stability and excellent flow characteristics. Heparin is linked to the Sepharose matrix by reductive amination which is a very stable binding, even in alkaline conditions. The stability of this medium is limited only by the heparin itself. The essential characteristics of Heparin Sepharose 6 Fast Flow are summarized in Table 1. 2 (A) (B) Figure 1. Heparin consists of alternating hexuronic acid (A) and D-glucosamine residues (B). The hexuronic acid can be either D-glucuronic acid (top) or its C-5 epimer, L-iduronic acid. R1= –H or –SO3 -; R2= –SO3 - or –COCH3. p. 3 p. 4 Table 1. Characte