Affinity Chromatography Solutions for Rapid Antibody and His-Tagged Protein Purification
Affinity chromatography (AC) is a crucial step for purifying biologics in life science laboratories and the biopharmaceutical industry. It is especially useful for single-step reduction of major contaminants from antibody and his-tagged protein samples, based on specific interactions with affinity ligands.
For screening, preparative purification and other research applications, Sartobind®️ IDA and our next-generation Rapid A membranes are available in a ready-for-use syringe filter format, which reduces workflow steps to minimize setup time and ensure simple, flexible, and highly selective purification.
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Affinity Chromatography and its Applications
Principle of Affinity Chromatography
When a complex mixture such as a cell culture supernatant is applied through the affinity chromatography membranes in Sartobind® Lab units, convective mass transport leads to efficient capture of the molecule of interest (MOI), while impurities such as host cell proteins (HCPs) and DNA flow through the membrane. A wash step may be used to increase purity before the MOI is eluted.
Need to concentrate or buffer exchange your sample before loading?
Antibody Purification
Protein A is a bacterial cell wall protein derived from the bacterium Staphylococcus aureus. It has a high affinity for the Fc region of immunoglobulin G antibodies, including mAbs.
When immobilized on a chromatography matrix, Protein A enables the selective capture of antibodies from complex mixtures, such as cell culture supernatants or serum samples. This method provides highly pure antibody samples that can be used in applications ranging from immunoassays to therapeutic antibody production.
His-Tagged Protein Purification
Divalent metal ions have an especially high affinity for histidine residues. In research laboratories, recombinant proteins are commonly engineered with a polyhistidine tag at the N- or C-terminus, facilitating their selective capture by immobilized metal affinity chromatography (IMAC).
Due to its reproducibility, selectivity and mild elution conditions, this method can simplify purification to a single-step process that produces samples of pure, native proteins, ready for structural and functional studies.
Affinity Chromatography Resources
Frequently Asked Questions
Affinity chromatography is based on specific interaction of a biomolecule with the chosen ligand. It can be used for capture purification of antibodies and other Fc containing molecules (Protein A) or proteins engineered with a his-tag (IDA).
Sartobind®️ IDA Lab can also be used for the capture or removal of divalent metal ions from environmental samples.
Sartobind® Lab is currently offered with Protein A or iminodiacetic acid (IDA) ligands.
Affinity chromatography is based on specific interaction of a biomolecule with the chosen ligand. It can be used for capture purification of antibodies (Protein A) or proteins engineered with a his-tag (IDA).
Sartobind® IDA Lab can also be used for the capture or removal of divalent metal ions from environmental samples.
One or four units, a pair of Luer to UNF adapters and a quick start guide.
Two units, a pair of Luer to UNF adapters and a quick start guide.
Yes, they are. While we have taken steps to minimize our environmental impact with quick start guides in the product packaging, if you need more information on how to use our products, the full instruction manual can be downloaded from the relevant product page in our eShop.
Both membrane chromatography units can be used with or without equipment. A syringe or peristaltic pump can be connected directly to the Luer lock inlet for fast purification with minimal setup time. Alternatively, a pair of adapters are included for compatibility with most liquid chromatography systems.
The bed volume is also referred to as the membrane volume (MV).
- Sartobind® Rapid A Lab: 0.5 mL
- Sartobind® IDA Lab: 2.1 mL
These chromatography units are stable in most common purification buffers. Oxidizing agents should be avoided.
Unlike with resin chromatography, there is no possibility of air bubbles disrupting Sartobind® membranes. This eliminates the need to degas buffers, saving considerable preparation time.
For optimal biomolecule capture, we recommend equilibration with a buffer similar in composition to the sample to be purified. This is a rapid process that adds only a few seconds to the purification cycle.
Although Sartobind® membranes are resistant to blocking, we recommend that all samples be pre-filtered before loading. To maximize productivity, a Minisart® syringe filter can be connected to the Sartobind® Lab inlet for simultaneous prefiltration and sample loading.
If the conditions for optimal capture of your molecule are unknown, we recommend screening with different metal ions loaded on the membrane. Suggested metal ions include nickel (Ni2+), cobalt (Co2+), copper (Cu2+) and zinc (Zn2+).
A scouting purification can also be performed to determine optimal washing and elution conditions. When purifying by centrifuge, syringe or pump, use a multi-step elution with buffers of gradually increasing imidazole concentration. If a liquid chromatography system is used, apply a linear elution gradient. An appropriate method (e.g. SDS PAGE) should then be used to analyze the flowthrough and eluent fractions and identify which conditions provide the required level of purity.
Each Sartobind® Lab unit can be reused many times with proper regeneration and storage. To maintain the binding capacity of Sartobind® IDA Lab units, regular stripping and reloading of the chelated metal ions is recommended.
We recommend using a separate Sartobind® Lab unit for each target biomolecule to eliminate the risk of carryover.
Sartobind® Rapid A Lab already has a high dynamic binding capacity of ≥17.5 mg per cycle.
To purify larger quantities, two units can be run in series. Alternatively, you can take full advantage of the high flow rates with a single unit, by purifying one sample with multiple, smaller loads over several cycles.
Yes, there are. Not only is Sartobind® Rapid A Lab, like all other Sartobind® Lab units, manufactured using 100% renewable energy sources, but the ability to re-use it for up to 100 cycles has the potential to reduce the number of consumables you need, thereby reducing research waste. In addition, unlike alternative protein A resin columns and membrane chromatography units, Sartobind® Rapid A Lab can be stored at room temperature, reducing the need for energy-intensive cooling.