LMWH Purification

Anion-exchange Chromatography Method

LMWH is used widely these years. The average molecular weight of LMWH used in the clinical application is less than 5000Da.

Compared with normal heparin, low molecular weight heparin has better antithrombotic effects with a lower risk of bleeding. LMWH also has the advantages of long half-life, clear dose-effect relationship, and high bioavailability. LMWH is widely used in the prevention and treatment of thromboembolic diseases, and it is effective and safe.

The extraction of LMWH is chemical degradation, physical degradation, enzymatic degradation. The production cost of enzymatic degradation is too high. Y- rays reaction in physical degradation is difficult to control. Therefore, The main production of LMWH is the chemical degradation method.

After chemical degradation, the LMWH product is a mixture of the fraction with molecular weights from a hundred to nearly ten thousand. This product does not meet the standard of Pharmacopoeia. Industrial production must have an effective separation purification media to separate fragments of a certain molecular weight range.

GALAK Chromatography Technology Co., Ltd provides an effective Anion-exchange Chromatography to separate LMWH. GALAK chromatography packing material Bettsep® D501 can apply in industrial LMWH production with the advantage of high dynamic adsorption capacity and super pressure resistance.

Bettsep ® D501 is high loads, weak anion exchange media, designed for the purification of proteins, peptides, and antibodies. It uses the polyester hydrophilic substrate, the average particle size is about 50 um. It has the advantages of excellent Dynamic binding capacity (DBC), extremely low non-specific adsorption, and excellent recovery rate. Bettsep ® D501 can operate at a high flow rate because of its extremely narrow particle size distribution and low backpressure.

Purification Result

LWMH purification
M. W. < 2000 2000-4000 2000-8000 Xa I Ia Potency Ratio
Solid Sample 4279 5.3 43.7 83.8 131.6 34.8 3.8

The molecular weight distribution of LMWH passed the European Pharmacopoeia. The recovery is over 70%.

Tips:

  1. The recovery rate is related to the molecular weight of the cracking solution.
  2. Xa and potency ratio are affected by molecular degradation.
  3. The operation of packing chromatography media affects the results a lot.

Character For Bettsep® D501

Items Character
Substrate Hydrophilic porous polymer microspheres
Average Particle Size 50 μm
Dynamic Binding Capacity ≥ 100 mg Lysozyme/ml resin
Ion-Exchange Capacity (IEC) ≥ 0.10 meq/ml resin
PH value 3-12
Clearance (CIP) 0.1 M NaOH solvent
Storage 20% ethanol aqueous solution

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