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To Affinity and Beyond: Speedy and Specific Biomolecule 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 interaction with a ligand.
For screening and research applications, Sartobind®️ Protein A and IDA membranes are available in a ready-for-use syringe filter format, which minimizes set up time and ensures simple, flexible, and highly selective purification.
Fast affinity purification for IgGs and mAbs
Ready to charge with the metal ion of your choice
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.
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.
Discover the full range of Sartorius lab chromatography consumables and kits.
Find the technical specifications and ordering information for protein A affinity chromatography.
Refer to the technical specifications and ordering information for metal affinity purification.
Affinity membrane chromatography is a separation technique that uses ligands to purify biomolecules based on specific binding characteristics. It works in a similar way to column chromatography, but the ligands are coupled to a stationary phase consisting of a macroporous membrane rather than microporous resins. This allows faster purification and higher yields of antibodies such as mAbs and IgGs, and isolation of proteins that have been engineered with a tag to assist the purification process.
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.
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).
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.
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