Heparin Superflow Plus

Product Code	37334
Bead Size	60-160µm
Agarose concentration	6%
Spacer Arm	5 atoms, hydrophilic
Flow rate	1500cm/h*
Binding Capacity	12mg AT3/ml resin**
Cleaning	0.5M NaOH, 3 hours
Storage Conditions	20% ethanol, 2-8C
Cycle Times	100
Temperature stability range	0-60 C
Functional ligand	Unfractionated Porcine heparin
Leaching	<0.1 ppm
pH stability range	2-13
Chemical stability	High salt, 0.1M NaOH, 6M guanidine, 70% ethanol, 50% Methanol
Drug master file	U.S. Food & Drug Administration

*measured on 1.5 X 5cm column
**Load = 10mg AT3


The Ligand
Heparin is a naturally occurring linear glycosaminoglycan consisting of a repeating dimer of a-L-dipyranuronic acid 2-sulfate and 2-deoxy-2-sulphamino-a-D-glucopyranose 6-sulfate. Heparin exists in a wide range of molecular weights from 5,000-30,000 Daltons. The primary linkages are 1-4 with very little 1-6 branching. Minute amounts of other sugars are also present. Heparin is highly charged and strongly acidic.

Binding and Elution
Immobilized heparin interacts with proteins by two different mechanisms: It can function as an affinity ligand, in which case the protein (such as a coagulation factor) is eluted with a buffer containing either salt or heparin. Heparin's anionic sulphate groups also give it the ability to function as a high-capacity cation exchanger. In this case, the protein is recovered by gradient elution with salt.

With heparin’s unique combination of affinity for many proteins and ion exchange, good purification factors can be achieved despite relatively small binding affinity differences between proteins.

No Detectable Leaching of Heparin
Heparin Actigel is made from our Actigel ALD activated resin. Actigel ALD creates exclusively uniform, stable secondary amine linkages between the reactive monoaldehyde groups and primary amino groups of the heparin.

The stability of the secondary amine is so high that leaching of the heparin is undetectable at 0.1 ppm, the sensitivity limit of the assay. Moreover, Actigel ALD does not release toxic leaving groups; the leaving group during coupling is water.

Easy Sanitization with NaOH
Traditional heparin resins are limited by the fact that they cannot be treated with high concentrations of NaOH, the standard sanitization method for ion exchange and gel filtration media.

In contrast, Sterogene’s heparin media can be sanitized using 0.5M NaOH. About 3-5 bed volumes are required. The resins can also be stored in 0.05M NaOH for extended periods. The stability of the coupled heparin with the stable secondary amine linkage allow these heparin gels to be used in the pH range of 2-13.

Highly Reproducible Resins
Irreproducible coupling is most often due to inactivation of the activated groups, a frequent problem with traditional coupling chemistries. Using ALD chemistry for heparin immobilization, coupling efficiency is high and very reproducible as no hydrolysis or inactivation of active groups take place during coupling in a side reaction. This provides a highly reproducible resin, well-suited for both research and manufacturing.

Purification Applications
Complement and coagulation factors
RNA and DNA polymerases
Restriction endonucleases
Ligases
Protein synthesis factors
Lipases and lipoproteins
Viral proteins
Drug Master File

Sterogene is a cGMP manufacturer of bioprocessing resins. Heparin Actigel is currently used in the production of an FDA-approved injectable therapeutic. A Drug Master File is on file with U.S. FDA.

Storage
The resins (as supplied) are stable for at least 2 years at 4oC. They should be stored between 4oC and 8oC at pH 7.0 with a preservative such as 0.02% merthiolate, 0.02% sodium azide or 20% ethanol. When packed into a column, Heparin gels can also be stored in 0.05M NaOH for extended periods of time (over 6 months).

Prepacked Columns and Kits:
Heparin Superflow Plus Prepacked Column