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Heparin Affinity Chromatography Medium

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  • Time of issue:2022-07-13 22:29
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(Summary description)

Heparin Affinity Chromatography Medium

(Summary description)

  • Categories:News
  • Author:
  • Origin:
  • Time of issue:2022-07-13 22:29
  • Views:
Information

Background

In 1916, under the guidance of Johns Hopkins University physiologist William Howell, second-year medical student Jay McLean extracted a highly sulfated polysaccharide from the liver and named it heparin. Heparin is a linear polymer, a mucopolysaccharide sulfate consisting of alternating glucosamine, L-iduronide, N-acetylglucosamine and D-glucuronic acid.

 

The affinity between heparin and biomolecules

The secondary amine groups in the glucosamine skeleton of heparin can form hydrogen bonds with specific biomolecular structures. At the same time, there are a large number of sulfonic acid groups and acetate groups in the heparin molecule, which makes heparin extremely negatively charged and can interact with other positively charged molecules. The groups of heparin form ionic bonds and these two modes of action make heparin have an affinity for a variety of proteins.

 

Glycosaminoglycan (GAG)-binding protein

Heparin molecules contain glycosaminoglycan (GAG) structures, and most of the chemical and physical properties of heparin are related to the structure or sequence, conformation, chain flexibility, molecular weight, and charge density of GAGs. There are more than 100 GAG-binding proteins reported in the literature. The binding of GAG to proteins has important effects on blood coagulation, lipid transport, cell growth and migration, and development. Through its own GAG structure and properties, heparin can be used for antithrombin, heparin cofactor II, t-plasminogen activator, fibroblast growth factor, endothelial cell growth factor, hepatocyte growth factor, and keratinocyte growth. Purification of factors, L-selectin, P-selectin, extracellular superoxide dismutase, lipoprotein lipase, fibronectin, laminin, and other proteins.

 

The GAG ​​structure of heparin can also be used as a binding receptor for some viruses. Using this affinity for viruses, heparin affinity chromatography media can be used to purify viruses such as adeno-associated virus, herpes virus, and pseudorabies virus. Heparin affinity chromatography has been widely used in the purification of AAV2.

 

Nucleic acid binding protein

Because heparin itself contains a large number of sulfonic acid groups and acetic acid groups, it is considered to be the substance with the highest negative charge density among the known biological macromolecules. The polyanionic structure of heparin can simulate the charged properties of nucleic acids, thereby producing affinity with nucleic acid-binding proteins. Therefore, heparin affinity chromatography can play a great role in the purification of nucleases and transcription factors.

 

The interaction between the above-mentioned several heparins and target proteins can be weakened by increasing the ionic strength. Therefore, when using heparin affinity chromatography medium for protein purification, the protein sample is usually placed in a solution of lower ionic strength, so that It can be well combined with the heparin medium, and then the target protein can be separated from the heparin medium by increasing the ionic strength.

 

Mars Heparin/Heparin Starose 6 Fast Flow

The Mars Heparin/Heparin Starose 6 Fast Flow medium was prepared by using the affinity of protein and heparin to couple heparin to the agarose matrix by covalent bonding, which can effectively capture the heparin-affinity in complex samples. protein, to achieve the separation and purification of specific proteins. At present, this kind of medium has been widely used in scientific research, molecular diagnosis, pharmaceutical and other biological fields.

 

Suzhou Nanotein provides different types and specifications of heparin affinity chromatography media, which can meet the needs of customers for various application scenarios such as different pressures/flow rates, resolutions, and loadings.

 

Product Name

Average Partical Size (μm)

Dynamic Binding Capacity (mL)

Pressure (MPa)

Maximum Speed (cm/h)

PH

Starose Mars Heparin

75

20 mg LZM

0.5

700

4-12

Heparin Starose 6 Fast Flow

90

25 mg LZM

0.3

600

4-12 (4-13)

Heparin Starose 6 High Performance

34

25 mg LZM

0.3

150

5-10

 

Case:

Process

Volume (ml)

Total Protein Concertration (mg/mL)

Total Protein Weight (mg)

Target Protein Concertration (mg/mL)

Target Protein Weight(mg)

Purity (%)

 Yield (%)

Sample

220

18.4

4048

15.2

3344

82.6

 

Flow

550

2.82

1554

1.5

834

53.7

 

Elute

160

15.5

2482

15.1

2408

97.2

72

 

 

Results: In the above application case, the recombinant hot-start DNA polymerase expressed in the E. coli system was purified using the first-generation heparin affinity medium Heparin Starose 6 Fast Flow of Nanotein. The purity of the final purified protein detected by HPLC was higher than 97%. The yield is greater than 70%.

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