IntroductionPall Heparin HyperD™ M composite chromatography sorbent is an affinity preparative sorbent designed for the purification of biological molecules that bind to heparin, such as coagulation factors, growth factors, lipoproteins... The sorbent provides high binding capacity at high flow rates. Heparin HyperD M unique composite structure was chosen to provide superior dynamic capacity at high linear velocities.
Heparin HyperD “gel-in-a-shell” sorbent is comprised of a porous rigid mineral bead containing heparin bound hydrogel filled pores. Heparin HyperD M has an average particle size of 80 µm and is used for preparative scale purification of ATIII. The sorbent can be packed in column sizes from mL to more than hundred liters and operated at high flow rates with low backpressure. Heparin leakage is minimized due to the stable chemical link of the heparin molecule to the sorbent.
Heparin HyperD M is available as ready-to-use labpacks suspended in 1 M sodium chloride with 20 % ethanol as bacteriostatic. Larger bulk quantities are also available upon request.
Features and Benefits
- High dynamic binding capacity at high linear flow rate
- Reference method for large-scale purification of Antithrombin III (AT III)
- Purification of various proteins from laboratory to production scale
Heparin HyperD™ M Affinity Sorbent ApplicationsHeparin is a mucopolysaccharide known for its anticoagulant and clarifying actions and is composed of equimolar quantities of glucosamine and glucuronic acid, alternatively linked by alpha-1,4 glycosic bonds.
A certain number of its hydroxyl groups are esterified with sulfuric acid, especially those on C-6 of glucosamine. Other groups are also sulfated, including C-3 of glucosamine and C-2 of glucuronic acid. The main characteristic of heparin is that it contains a large number of amino groups combined with sulfate groups, the latter being quite labile in acidic medium.
The molecule contains small quantities of other sugar, such as galactose and xylose, and amino acids. As a result of its composition and its biochemical role, heparin has the property to combine with a number of proteins, enzymes and in general with polycationic organic compounds. It is also combined with alkaloids, antibiotics, stains and hormones.
Main Applications Include
- Reference method for large-scale purification of antithrombin III (ATIII)
- Other coagulation factors such as Factor IX, Factor VII, Factor XI, Factor XII and XIIa
- Lipoprotein lipases purification
- Lipoproteins (LDL, VLDL, VLDL apoprotein, HDL)
- Growth hormones
- Growth factors: FGF (fibroblast growth factor), ECGF
- DNA- and RNA-related enzymes
- Purification of various other enzymes (collagenase, α-L-iduronidase, hyaluronidase and lysozyme), fibronectin, fibronectin fragments and hormones receptors
Main Properties of Heparin HyperD M Affinity Sorbent
|Particle Size||80 µm (av.)|
|Dynamic Binding Capacity for hu ATIII (600 cm/h)||> 25 mg/mL|
|Recommended Operating pH Range||3 – 13|
|Volume Changes due to pH and Ionic Strength||Non compressible|
|Pressure Resistance||70 bar (1,000 psi)|
Chemical and Mechanical StabilityThe pH stability is the same as for the free soluble heparin: between 3 and 13. Dissociating agents and detergents have generally no effect on heparin sorbent. Heparin HyperD M can be cleaned with sodium hydroxide in concentrations of 0.01 to 0.1 M.
The non compressible HyperD matrix can withstand very high flow rates without any risk of bed collapse. As a result, Heparin HyperD M saves user time and preserves the biological integrity of the purified proteins. The mechanical properties of Heparin HyperD M sorbent remain constant across a wide range of velocities. Minimum pressure drop, even at high linear velocity, assures direct, predictable scale up to any volume (see Figure 1).
Figure 1: Pressure vs. Linear Flow Velocity
CapacityHeparin HyperD M maintains high binding capacity, even at high linear velocity. It is commonly used at large scale for the production of pharmaceutical grade ATIII. Production scale columns (>100 L) can be operated at high linear velocities (> 200 cm/h) while maintaining capacity with minimal backpressure (Figure 1). Its capacity for ATIII is higher than 25 UI/mL even at 600 cm/h with a 10 cm bed height (Figure 2).
Figure 2 : Dynamic Binding Capacity vs. Linear Velocity
ValidationThe heparin used for the production of Heparin HyperD M has a North American origin and is from porcine intestinal mucosa.
The heparin is produced in compliance with the applicable requirements of the FDA’s Good Laboratory Practices and Good Manufacturing Practices regulations.
A validation file can be provided to industrial customers to support the regulatory requirements for producing clinical and approved therapeutics
Sorbent Ordering Information
|Size||Heparin HyperD Affinity Sorbent|