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 Chromatography Process

HyperD and Trisacryl affinity resins are designed for capturing proteins from cell culture supernatants and bacterial lysates.  
 

Attributes 

Heparin HyperD 

Lysine HyperD 

Blue Trisacryl 

Particle size 

80 µm 

70 µm 

40 – 80 µm 

Dynamic Binding Capacity (10 % BT) 

> 25 mg/mL 

> 10 mg/mL 

Operating pH 

3 – 13 

3 - 13 

4 - 10 

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Sartorius offers membranes for affinity chromatography for research and process development scale. These membrane adsorbers are ready to use and save you time. Switch to single-use chromatography and eliminate packing, cleaning and revalidation steps. 

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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. 

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Introduction

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 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

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

Chemical and Mechanical Stability

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

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. 

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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
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)

Lysine HyperD Affinity sorbent employs a high capacity hydrogel polymerized within the large pores of a rigid bead.

This design combines the desirable characteristics of a soft, high capacity gel with the dimensional stability of a rigid bead.

Dynamic binding capacity is virtually independent of linear velocity. High productivity is obtained as a result of this combination. The sorbent packing is fast due to the high density of the beads.

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. 

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Introduction

Blue Trisacryl M dye-affinity chromatography sorbent is used for the purification of a wide variety of enzymes and proteins such as kinases, interferons, and some coagulation factors. It is also used for albumin purification or albumin depletion in samples for proteomics research.

Blue Trisacryl M sorbent is based on Trisacryl support, a macroporous non ionic synthetic polymer on which blue dye is covalently immobilized through a stable six-carbon spacer arm to prevent leakage of the dye in normal working conditions.


Features and Benefits

  • Unique multi-modal interaction mechanism
  • Strong bond of dye-to-sorbent prevents leakage of the dye
  • Multiple laboratory to production scale applications


Presentation and Storage

Blue Trisacryl M sorbent is available as ready-to-use labpacks suspended in 1 M sodium chloride with 20% (v/v) ethanol as bacteriostatic, or in drums for process-scale 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

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

Chemical and Mechanical Stability

The chemical stability of Blue Trisacryl M sorbent is a function of the synthetic nature of Trisacryl matrix and the enhanced stability of the ligand coupling mechanism. The sorbent can be operated up to 3 bar backpressure while maintaining good flow rate properties.

Capacity

The binding capacity of Blue Trisacryl M sorbent depends on the protein, the composition of the feedstock and parameters such as pH. Note that the capacity for a given protein may differ according to the animal species (for example, the binding capacity for bovine albumin is lower than for human albumin).

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 Sartbind 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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Rapid Cycling Chromatography in mAb Capture

It takes 3 to RCC – Learn more about our membrane platform enabling highly productive Rapid Cycling Chromatography for intensified mAb capture.

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