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Affinity Chromatography

Affinity chromatography is used to capture molecules by specific interaction between the chromatography ligand and the target. This allows the removal of major contaminants to a low concentration in a single process step. Affinity chromatography is frequently used for downstream purification of proteins & vaccines. Sartorius offers a broad selection of resins and membranes for this purpose. 

Find the Right Solution for Your Affinity Chromatography Process

Resins
Attributes Ceramic HyperD Q Ceramic HyperD DEAE Ceramic HyperD CM Hypercel Star AX 
Particle diameter 50 µm  50 µm  50 µm  80 µm 
Dynamic Binding Capacity (10 % BT)  > 85 mg/mL BSA  > 85 mg/mL BSA  > 60 mg/mL IgG  > 100 mg/mL BSA 
Functionality Strong Anion Exchange  Weak Anion Exchange Weak Cation Exchange Weak Anion Exchange (Salt tolerant)
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Membranes

Affinity Resins

Heparin HyperD

Heparin HyperD resin is suited for the purification of heparin binding proteins like coagulation factors, growth hormones, lipoproteins, and DNA/RNA processing enzymes. 

Description

Applications

Heparin HyperD™ M Affinity Sorbent Applications

Heparin 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

Specifications

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
Ligand Porcine heparin
Recommended Operating pH Range 3 – 13
Volume Changes due to pH and Ionic Strength Non compressible
Pressure Resistance 70 bar (1,000 psi)

*Capacity determined using hu ATIII at 72.5 UI/ml in 20 mM Tris-HCl, 0.3 M NaCl, pH 7.4. Elution with 20 mM Tris-HCl, 2 M NaCl, pH 7.4 at 600 cm/h, 10 cm bed height

Performance

The 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.

Capacity

Heparin 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. Its capacity for ATIII is higher than 25 UI/mL even at 600 cm/h with a 10 cm bed height.

Validation

The 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

Ordering Information

Heparin HyperD Affinity Sorbent
Description Part Number
25 mL 20029-039 
100 mL  20029-021 
1 L 20029-013
10 L 20029-054
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Lysine HyperD

Lysine HyperD affinity resin is used for the purification of lysine binding proteins like plasminogen. It provides high protein binding capacities at high flow rates. 

 

Description

Introduction

Lysine HyperD™ sorbent is a high-speed, high-capacity affinity preparative sorbent for the purification of biological molecules that bind to lysine, such as plasminogen from human or animal species plasma. The sorbent provides high binding capacity at high flow rates.

Features and Benefits

  • High dynamic capacity
  • Ridgid sorbent provides high flow rates

Specifications

Particle size   70 µm (av.)
Ligand L-Lysine
Recommended Operating pH Range 3 – 13
Volume Changes due to pH and Ionic Strength Non compressible
Pressure Resistance 70 bar (1,000 psi)

Performance

Ordering Information

Lysine HyperD Affinity Sorbent
Description Part Number
25 mL 20059-036
100 mL 20059-028
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Blue Trisacryl

Blue Trisacryl is an affinity resin that is used for the purification of enzymes, interferones, and some coagulation factors as well as for the removal of albumin. The resin uses Trisacryl based bead which carries the blue dye ligand. 

 

Description

Applications

Blue Trisacryl M Sorbent can be Used for the Purification of Many Proteins

Blue Trisacryl M sorbent is a group-specific adsorbent with affinity for a wide variety of enzymes. Some proteins interact biospecifically with the dye due to its structural similarity with nucleotide cofactors (ADP, NADP). Other proteins like albumin or interferon bind by a combination of hydrophobic, electrostatic and pi-pi interactions.

Applications include :

  • Enzymes which need NAD as cofactor (kinases, dehydrogenases, phosphatases…)
  • Other enzymes (sulfatases, RNA polymerases, mono-oxygenases, oxydoreductases)
  • Albumins from plasma
  • Transferring from plasma
  • _α1-acid glycoprotein, α-fetoprotein, α1-proteinase inhibitor
  • Serine proteases
  • Interferons
  • DNA antibodies
  • Recombinant vaccines purification processes

Specifications

Main Properties of Blue Trisacryl M Sorbent

Particle size 40 – 80 μm
Ligand Blue dye
Exclusion limit 107 Da
Capacity for human albumin1 ≥10 mg/mL
Thermal stability 2 to 121 °C
Working pH 4 to 10
Cleaning pH 1 to 12 (short term)
Working pressure at 100 cm/hr Up to 3 bar (45 psi)

1 Determined in PBS buffer using 5 mg/mL protein, column dimensions 16 mm I.D. x 6 cm bed height, flow rate 25 cm/hr

Performance

Ordering Information

Blue Trisacryl M Sorbent
Description Part Number
25 mL 25896-045
100 mL 25896-010
1 L 25896-028
5 L 25896-044
10 L 25896-036
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Affinity Membranes

Sartobind® SC

The benefits of high throughput membrane adsorbers such as reducing time and buffers can now be experienced using the Sartobind SC membrane. Ideal for capture of impurities such as host cell proteins and DNA, this membrane is available in nano sizes and well suited for replacing columns in process development and research use.  

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