Affinity Chromatography
Affinity chromatography is a powerful technique that continues to revolutionize the purification of proteins and other biomolecules in biopharmaceutical manufacturing. By leveraging the specific interactions between target molecules and immobilized ligands, this method ensures high selectivity and efficiency in isolating the desired target molecule from complex mixtures and significantly reduces processing volumes in following unit operations.
Affinity Chromatography Portfolio
Find the Right Solution to Selectively Purify Your Target Molecule
Discover how Sartorius’ diverse affinity chromatography media and ligand combinations can help streamline your downstream process.
Affinity chromatography membranes combine the high selectivity of affinity ligands with the productivity of membrane-based chromatography matrices. With convection as the dominant mass transport mechanism, target molecules are quickly transported to the binding site. The larger pore sizes of chromatography membranes enable higher flow rates and greater productivity, making them especially well-suited for processing larger molecules such as virus particles.
| Sartobind® Rapid A | Sartobind Convec® SC |
|---|---|---|
Ligand | CEX, HIC | AEX, HIC |
| Base Matrix | Agarose | Cellulose |
| Interaction mechanism | Fc-region affinity | Heparin-like pseudo-affinity | CEX |
| Bed-height | 4 mm | 8 mm |
| Focus applications | mAb capture | Capture of traditional vaccines and VLPs (e.g., influenza, rabies, and encephalitis) |
Affinity chromatography resins have a long legacy in purification of biopharmaceuticals. The extended interaction area created by the the porous structure inside the resins beads, leads to high-binding capacities and makes this material a common choice for commercial manufacturing set-ups.
| Heparin HyperD® | Lysine HyperD® | Blue Trisacryl® | |
|---|---|---|---|
| Ligand | Heparin | L-Lysine | Blue dye |
| Base Matrix | 80 µm silica beads | 80 µm silica beads | 40 – 80 μm macroporous non-ionic synthetic polymers bead |
| Interaction mechanism | Heparin Affinity | Lysine Affinity | Blue dye affinity |
| Focus Applications | Heparin binding proteins (e.g., coagulation factors, growth hormones, lipoproteins, and DNA/RNA processing enzymes.) | Lysine-binding protein (e.g., plasminogen (human and animal)) |
|
| View Products | View Products | View Products |
Monoliths
CIMmultus® Oligo dT – Affinity for RNA
Oligo dT ligands covalently bound to solid support
Hybridizes to poly-adenylated tail found on most eukaryotic mRNAs, or synthetized onto the molecule during IVT
Fast and efficient capture and purification of mRNA with a poly-adenylated tail, from various sources
Selectively removes impurities from samples
Convenient approach for initial purification of mRNA
Available with:
Supporting Products & Services
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Highlight Assets
Mastering Affinity Chromatography for Biopharma Excellence
The term “affinity chromatography” was coined in 1968 by Cuatrecasas, Anfinsen, and Wilchek, though the technique’s roots date back to the early 20th century with the advent of liquid column chromatography. Thanks to its simplicity and selectivity, affinity chromatography has evolved far beyond its early analytical use in research and development. Today, preparative affinity chromatography is a cornerstone of biopharmaceutical manufacturing.
Sartorius offers a broad portfolio of media: membranes, resins, and monoliths - to deliver customized affinity solutions tailored to a wide range of biomolecules. By aligning purification strategies with the specific needs of each molecule, Sartorius continues to support advancements in drug development and production—supporting success in biopharmaceutical manufacturing.
Frequently Asked Questions
Affinity chromatography is a technique used to separate and purify molecules based on specific interactions between the target molecule and a ligand attached to the stationary phase. The feed stream passes through a chromatographic media containing the ligand, which selectively binds the target molecule. Unbound substances are washed away, and the target molecule is eluted by altering conditions, such as pH or ionic strength, to disrupt the interaction. This method is highly effective for isolating proteins, enzymes, and other biomolecules with high specificity and purity.
Generally, affinity ligands can be divided into biological ligands (such as antibodies, Protein A, and enzymes) or non-biological ligands (such as IMAC, polymers, and dyes).
Affinity chromatography can purify a wide range of molecules, including proteins, enzymes, antibodies, nucleic acids, and glycoproteins. It is particularly effective for isolating specific proteins or peptides from complex mixtures due to the high specificity of the ligand-target interactions. Additionally, affinity chromatography can be used to purify tagged proteins, such as those with histidine tags, and other biomolecules that have a known affinity for a particular ligand. This versatility makes it a valuable tool in biochemistry and molecular biology for obtaining high-purity samples.
Depending on the type of affinity ligand, at Sartorius affinity chromatography consumables are available either as membranes, resins, or monoliths.
