Shop All Affinity Columns, Media, & Standards

CaptureSelect™ LC-lambda (Hu) Affinity Matrix (Thermo Scientific™)

This is one of a group of CaptureSelect™ Antibody Toolbox™ affinity resins designed for the simple, single-step purification of all antibody subtypes and fragments. Unlike the common bacterial coat protein matrices (e.g., Protein A, G, and L), the Antibody Toolbox resins were specifically designed to purify any antibody format with high affinity and specificity. These unique affinity ligands are immobilized on a high capacity agarose support for lower-pressure antibody purification applications.

CaptureSelect™ Antithrombin III Affinity Matrix (Thermo Scientific™)

This is one of a group of CaptureSelect™ protein purification affinity resins developed for the simple, single-step purification of a variety human plasma and serum proteins. This unique selection of affinity purification resins can purify proteins or other non-antibody biomolecules from either recombinant or native sources with a high degree of affinity and specificity, and are immobilized on a high capacity agarose support.

POROS™ CaptureSelect™ CH1-XL Affinity HPLC Column, 4.6 x 50 mm (Thermo Scientific™)

POROS CaptureSelect CH1-XL Affinity HPLC columns incorporate the advantages of high-speed POROS chromatography resins with the unique selectivity of CaptureSelect affinity ligands. CaptureSelect CH1-XL Affinity HPLC columns recognize the CH1 domain of human IgG antibodies, enabling quantitation and small-scale purification of all IgG subclasses (IgG1, IgG2, IgG3, and IgG4), as well as Fab fragments and fusion proteins containing the intact CH1 domain, independent of light-chain iso-type and source material.Analytical groups supporting biologic drug development require rapid, sensitive, and precise methods for protein quantitation from cell culture, fermentation harvests, or purification intermediates. It is critical for biopharmaceutical manufacturers to be able to monitor bioreactor product titers to determine when to harvest or abort a reactor cycle.

Features of POROS CaptureSelect CH1-XL Affinity HPLC columns include:
Versatile—applicable for IgG Fab fragments, all human IgG subclasses, and IgG fusion proteins with intact CH1 domain
Specific—no cross-binding to bovine IgG
Fast—POROS resins enable quantitation or purification in minutes
Hands-off operation—HPLC method can be automated for a streamlined operation
Robust—designed to deliver high-performance purification over hundreds of chromatography cycles

High speed with high specificity
POROS high performance chromatography allows for the quantitation of product from clarified harvest or partially purified intermediates—typically in under three minutes. Moreover, only minimal sample preparation (0.2 µm filtration) is required, and the eluted product is highly purified and suitable for further analysis, including mass spectrometry.

Automation
POROS Affinity HPLC columns are for use with HPLC systems, which allows the automation of chromatography methods, data collection, and data analysis. In addition, the immobilized affinity ligand enables one method and buffer system to be used across a class of molecules, streamlining analytical approaches.

Multiple cycles
Proper sample preparation, adherence to column operating instructions, and the use of an optimized column cleaning regimen results in strong re-use performance. POROS Affinity HPLC columns can be cycled hundreds of times for cost-effective and reproducible quantitation or purification.

Main characteristics

Protein binding capacity: 26.2 mg/mL for human IgG; 8.8 mg/ml human Fab fragment
Species specificity: human
Column material: PEEK polymer
Frit: 2 µm, titanium
POROS resin: cross-linked poly(styrene-divinylbenzene)
CaptureSelect ligands: single-domain monospecific antibody fragment produced in Saccharomyces cerevisiae

HisPur™ Cobalt Superflow Agarose (Thermo Scientific™)

Thermo Scientific HisPur Cobalt Superflow Agarose is a tetradentate-chelating Superflow 6 support charged with divalent cobalt, designed for high-purity, large-scale purification of polyhistidine-tagged proteins.

Features of HisPur Cobalt Superflow Agarose:

Robust—highly crosslinked 6% agarose Superflow beaded support tolerates linear flow rates up to 1200 cm/hour with minimal leaching of cobalt due to a tetradentate chelator
High yields—binds 20 mg of 6xHis-GFP per mL of resin at a linear flow rate of 150 cm/hour, can bind greater than 30 mg per mL of resin depending on flow rate
High purity—specificity of cobalt binding to histidine-tag generally results in elution fractions with less than 10% contamination of non-specific proteins
Compatible—maintains function after exposure to a wide variety of chemicals and pH values
Cost effective—resin is stable through multiple cycles of cleaning and reuse

HisPur Cobalt Superflow Agarose is a highly crosslinked, durable resin that does not compress at flow rates used for medium- to large-scale FPLC purifications. The resin holds up well to a variety of chemicals and pH values, and is compatible with common clean-in-place procedures. Compared to nickel and other ligands commonly used for immobilized metal affinity chromatography (IMAC), cobalt provides greater specificity for His-tagged protein purification. The specific binding of HisPur Cobalt Superflow Agarose results in purer elution fractions, reducing the need for extra polishing steps.

Applications:
• Large scale FPLC purification of polyhistidine tagged proteins

Affinity chromatography is often used as a quick and easy approach for the purification of recombinant proteins. One such method of purification involves the binding of a polyhistidine tag to a divalent metal cation that has been coordinated by a chelator immobilized on a beaded support. For immobilized metal affinity chromatography (IMAC) purification of His-tagged proteins, the type of bead, chelator, and metal immobilized influences purity, yield, and flow rate performance of these purifications.

The highly crosslinked 6% agarose Superflow beaded support tolerates larger linear flow rates than agarose resins that have lower amounts of crosslinking. The HisPur Cobalt Superflow Agarose protocol utilizes gentle wash and elution conditions and typically produces greater than 90% pure target protein. Protein purities achieved with HisPur Cobalt Superflow Agarose are generally higher than those achieved with nickel IMAC resins making HisPur Cobalt Superflow Agarose a valuable tool for users interested in one-step purifications. Additionally, HisPur Cobalt Superflow Agarose is perfect for larger scale purifications that require the resin to withstand medium pressures without compressing.

More Product Data
Performance characterization of HisPur Cobalt Superflow Agarose

Related Products
HisPur™ Cobalt Resin

Pierce™ High Capacity NeutrAvidin™ Agarose (Thermo Scientific™)

Thermo Scientific High Capacity NeutrAvidin Agarose is a beaded agarose resin of immobilized NeutrAvidin Protein, a modified form of avidin with exceptional biotin-binding characteristics for affinity purification methods.

Features of High Capacity NeutrAvidin Agarose:

NeutrAvidin protein—purified, deglycosylated avidin protein (60kDa, pI 6.3); tetrameric with four biotin-binding site per molecule
Agarose resin—support is crosslinked 6% beaded agarose (CL-6B), the most popular resin for protein affinity purification methods
Inert and stable—NeutrAvidin protein is immobilized by charge-free, leach-resistant covalent bonds, resulting in low nonspecific binding and enabling multiple uses without decline in yield
High capacity—this variety of beads has a dense load of immobilized NeutrAvidin protein, providing a binding capacity greater than 8 mg of biotinylated BSA per mL of resin

High Capacity NeutrAvidin Agarose consists of a specially purified and deglycosylated form of avidin that has been covalently immobilized at a high density onto high-quality crosslinked 6% beaded agarose. Compared to native avidin, NeutrAvidin protein is more neutral and exhibits much less nonspecific binding properties in biotin-binding applications. The agarose beads have physical and chemical properties that enable them to be used in a variety of batch- or column-type affinity procedures. Generally, NeutrAvidin products function as well or better than their streptavidin counterparts and are less expensive.

NeutrAvidin protein is deglycosylated native avidin from egg whites. Removal of the excess carbohydrate by an exclusive process yields a protein with a more neutral isoelectric point and less nonspecific binding properties. NeutrAvidin resins are prepared by covalently coupling the protein using efficient and stable chemistries, resulting in supports that are resistant to leaching and stable at pH 2-11. The products are excellent choices for a variety of small- or large-scale affinity purification applications involving biotinylated macromolecules, including separation of biotinylated molecules from samples and immunoprecipitation of antigens using biotin-labeled antibodies.

Properties of crosslinked 6% beaded agarose (CL-6B):
• Support pH Stability: 2 to 14 (short term); 3 to 13 (long term)
• Average Particle Size: 45 to 165 microns
• Exclusion Limit: 10,000 to 4,000,000 daltons
• Maximum Volumetric Flow Rate: approx. 1 mL/minute (for 1 cm diameter column)
• Maximum Linear Velocity: 30 cm per hour
• Maximum Pressure: less than 25psi (1.5 bar)

Related Products
Pierce™ NeutrAvidin™ Agarose
Pierce™ NeutrAvidin™ UltraLink™ Resin
Pierce™ NeutrAvidin™ Plus UltraLink™ Resin

Pierce™ Streptavidin Plus UltraLink™ Resin (Thermo Scientific™)

Thermo Scientific Pierce Streptavidin Plus UltraLink Resin is a beaded polyacrylamide resin of immobilized recombinant streptavidin protein for use in a variety of biotin-binding affinity purification methods.

Features of Streptavidin Plus UltraLink Resin:

Streptavidin—purified recombinant streptavidin protein (53kDa, near-neutral pI); tetrameric with four biotin-binding site per molecule
UltraLink Resin—acrylamide-based support provides an alternative to crosslinked beaded agarose supports for medium-pressure applications
High capacity—this variety of beads has a dense load of immobilized streptavidin, providing a binding capacity of at least 3 mg of biotinylated BSA per mL of resin

Applications:
• Purification of membrane antigens in conjunction with biotinylated monoclonal antibodies
• Isolation of cell-surface proteins that have been labeled with a cleavable biotinylation reagent
• Isolation of cell-surface glycoproteins using biotinylated Concanavalin A
• Recovery of single-stranded biotinylated DNA for dideoxy sequencing
• Streptavidin-coated plates used for direct solid-phase sequencing
In vitro mutagenesis with streptavidin-coated plates
• DNA fingerprinting with streptavidin-coated plates

Properties of Thermo Scientific UltraLink Resin:
• Support pH Stability: 1 to 13
• Average Particle Size: 50 to 80 microns
• Exclusion Limit: greater than 2,000,000 daltons
• Average Surface Area: greater than 250m2 per gram of beads
• Average Pore Volume: greater than 1.2 mL per gram of beads (>60% of bead volume)
• Pore Size: 500 to 1000 angstroms (50 to 100nm)
• Maximum Linear Velocity: 3000 cm per hour
• Maximum Pressure: 100psi (6.9 bar)

Pierce Streptavidin Plus UltraLink Resin consists of purified recombinant streptavidin that has been covalently immobilized at high density onto a beaded acrylamide support. Streptavidin is a tetrameric biotin-binding protein. UltraLink beads are hydrophilic, charge-free, highly crosslinked, rigid and durable. These properties make the affinity resin suitable for a variety of batch- or column-type affinity procedures that require medium-pressure chromatography. The resin provides an alternative to agarose supports when situations demand it. Biotinylated antibodies, proteins, peptides, nucleic acids and other molecules or interaction complexes can be captured, immunoprecipitated, removed or purified from samples using this streptavidin resin.

Streptavidin is a tetrameric protein containing four biotin-binding sites similar to avidin. The native protein from Streptomyces avidinii is carbohydrate-free, has an acidic pI of 5.5 and a mass of 75 kDa. The recombinant protein used to manufacture Thermo Scientific Pierce Streptavidin Resins has a near-neutral isoelectric point and a mass of 53kDa. Streptavidin generally has less nonspecific binding than avidin (from chicken egg white) because of the absence of carbohydrates.

UltraLink Resin is an azlactone-activated polyacrylamide support that is hydrophilic, charge-free, high capacity, highly crosslinked, rigid and porous. This support is especially useful for medium pressure techniques when using large sample volumes requiring fast-flow techniques (FPLC) and large-scale applications. Unlike typical beaded agarose supports, UltraLink Support does not crush or collapse when subjected to high flow rates that result in pressures exceeding 25psi.

Related Products
Pierce™ Streptavidin UltraLink™ Resin
Pierce™ Streptavidin Agarose
Pierce™ High Capacity Streptavidin Agarose

POROS™ CaptureSelect™ CH1-XL Affinity HPLC Column, 4.6 x 100 mm (Thermo Scientific™)

POROS CaptureSelect CH1-XL Affinity HPLC columns incorporate the advantages of high-speed POROS chromatography resins with the unique selectivity of CaptureSelect affinity ligands. CaptureSelect CH1-XL Affinity HPLC columns recognize the CH1 domain of human IgG antibodies, enabling quantitation and small-scale purification of all IgG subclasses (IgG1, IgG2, IgG3, and IgG4), as well as Fab fragments and fusion proteins containing the intact CH1 domain, independent of light-chain iso-type and source material.Analytical groups supporting biologic drug development require rapid, sensitive, and precise methods for protein quantitation from cell culture, fermentation harvests, or purification intermediates. It is critical for biopharmaceutical manufacturers to be able to monitor bioreactor product titers to determine when to harvest or abort a reactor cycle.

Features of POROS CaptureSelect CH1-XL Affinity HPLC columns include:
Versatile—applicable for IgG Fab fragments, all human IgG subclasses, and IgG fusion proteins with intact CH1 domain
Specific—no cross-binding to bovine IgG
Fast—POROS resins enable quantitation or purification in minutes
Hands-off operation—HPLC method can be automated for a streamlined operation
Robust—designed to deliver high-performance purification over hundreds of chromatography cycles

High speed with high specificity
POROS high performance chromatography allows for the quantitation of product from clarified harvest or partially purified intermediates—typically in under three minutes. Moreover, only minimal sample preparation (0.2 µm filtration) is required, and the eluted product is highly purified and suitable for further analysis, including mass spectrometry.

Automation
POROS Affinity HPLC columns are for use with HPLC systems, which allows the automation of chromatography methods, data collection, and data analysis. In addition, the immobilized affinity ligand enables one method and buffer system to be used across a class of molecules, streamlining analytical approaches.

Multiple cycles
Proper sample preparation, adherence to column operating instructions, and the use of an optimized column cleaning regimen results in strong re-use performance. POROS Affinity HPLC columns can be cycled hundreds of times for cost-effective and reproducible quantitation or purification.

Main characteristics

Protein binding capacity: 26.2 mg/mL for human IgG; 8.8 mg/ml human Fab fragment
Species specificity: human
Column material: PEEK polymer
Frit: 2 µm, titanium
POROS resin: cross-linked poly(styrene-divinylbenzene)
CaptureSelect ligands: single-domain monospecific antibody fragment produced in Saccharomyces cerevisiae

POROS™ CaptureSelect™ FSH Affinity Column, 4.6 x 50 mm (Thermo Scientific™)

Analytical teams that support biologic drug development require rapid, sensitive, and precise methods for protein quantitation from cell culture, fermentations broths, or purification intermediates. It is critical for biopharmaceutical manufacturers to be able to monitor bioreactor product titers to determine when to harvest or abort a reactor cycle. POROS™ CaptureSelect™ FSH HPLC columns incorporate the advantages of high-speed POROS™ chromatography resins with the unique selectivity of CaptureSelect™ ligands, enabling 3-minute quantitation of human follicle stimulating hormone (FSH).

Features of this class of chromotography columns:

• Broad utility—POROS™ A, G, and CaptureSelect™ affinity ligands address most biological drug classes
• Rapid—POROS™ resins offer fast quantitation or purification of immunoglobulins and fusion proteins
• Hands-off operation—HPLC method can be automated for streamlined operation
• Robust—delivers high-performance purification over hundreds of chromatography cycles

High Speed with High Specificity
The combination of CaptureSelect™ FSH affinity ligand with POROS™ 20 µm resin provides a high flow-rate, high performance chromatography solution for the rapid quantitation and small-scale purification of FSH.

POROS™ high performance chromatography allows for the quantitation of product from clarified harvest or partially purified intermediates—typically in less than 3 minutes. Moreover, only minimal sample preparation (0.2 µm filtration) is required, and the eluted product is highly purified and suitable for further analysis, including mass spectrometry.

Automation
POROS™ affinity chromatography columns are suitable for use with HPLC systems, allowing for the automation of chromatography, data collection, and data analysis. In addition, the immobilized affinity ligands enable one method and buffer system to be used across a class of molecules, streamlining analytical approaches.

Multiple Cycles
Proper sample preparation, adherence to column operating instructions, and the use of an optimized column cleaning regime results in strong re-use performance. POROS™ affinity chromatography columns can be cycled hundreds of times for cost-effective and reproducible quantitation or purification.

CaptureSelect™ Albumin RoboColumn (Thermo Scientific™)

CaptureSelect™ Human Albumin Affinity Matrix, available here in RoboColumn™ format, has been specifically designed for the purification of human albumin and albumin fusion proteins. RoboColumns are small chromatography columns used for fully automated and parallel chromatographic separations on robotic liquid handling workstations.

Features of this affinity matrix include:

• Highly selective for human albumin
• Unique elution characteristics allowing for gentle elution of albumin fusion proteins
• Excellent scalability
• Non-animal-derived

The high selectivity and yields obtained using CaptureSelect Human Albumin Affinity Matrix enable a robust and efficient purification process with excellent purity obtained in one step. The mild elution conditions make the resin suitable for the purification of albumin fusion proteins, especially those that are pH sensitive.

Free of animal components
Our CaptureSelect products are affinity ligands created by a proprietary technology based on camelid-derived single domain antibody fragments. The ligand is a 13 kDa single domain fragment comprising the 3 CDRs that form the antigen binding domain, efficiently produced by the yeast Saccharomyces cerevisiae in a production process free of animal components.

Characteristics:

Matrix: agarose-based, aldehyde-activated
Average particle size: 65 ± 10 µm
Ligand: CaptureSelect HSA affinity ligand
Ligand coupling method: aldehyde coupling via NH2 residues of the ligand
Binding capacity: ~15 mg albumin/mL resin
Elution conditions (neutral pH): 20 mM Tris, pH 7.4 with 2 M MgCl2 or with 1 M NaCl + 50%(v/v) propylene glycol or with 1 M NaCl + 0.5 M L-arginine
Elution conditions (acidic pH): 0.1 M glycine, pH 3.0
Formulation buffer: 20% (v/v) ethanol

RoboColumn is a registered trademark of ATOLL GmbH.

POROS™ EP 20 µm Column, 4.6 x 100 mm, 1.7 mL (Thermo Scientific™)

Applied Biosystems™ POROS™ Prepacked Immobilized Epoxide Affinity Column. POROS™ EP is based on an epoxide functional group designed for the immobilization of proteins through free amine groups.
20 micron particle size is used for high resolution and small scale preparative to semi-preparative separation of biomolecules.

POROS™ High Performance Chromatography™ Media and Pre-Packed Columns

A high performance chromatography resins for analytical to process scale separations.• Higher productivity: High throughput and high dynamic capacity associated with High Performance Chromatography™
• Chemical stability: Allows aggressive cleaning and sanitization
• Enhanced biomolecule access: Provided via large pores, ranging between 500-10000 Å
• Polystyrenedivinylbenzene particles: Yield a robust, easily packable matrix
• Develop better separations methods in a shorter time frame: The speed of High Performance Chromatography technology reduces weeks of experimentation to only a few hours of work, so you have plenty of time to explore all the variables of your separation

Reduce time-consuming sample prep

You can replace many sample preparation steps with high speed High Performance Chromatography technology. For instance, dialysis of large volumes of material can be replaced by high flow rate processing of the dilute sample. Elution in a small volume of new buffer accomplishes both buffer exchange and concentration.

Create novel assays for more efficient analysis

High Performance Chromatography technology is not limited to standard modes of separation. Both enzymes and affinity ligands can be immobilized on POROS media. By combining rapid on-column protein digestions with rapid on-column immunoassays and chromatographic separations, you can create entirely new assays with unlimited potential.

High capacity, high resolution, high speed
In contrast to conventional chromatography media, POROS™ High Performance Chromatography™ resins particles, are engineered to have two discreet classes of pores. Large "throughpores" allow convection flow to occur through the particles themselves, quickly carrying sample molecules to short "diffusive" pores inside. By reducing the distance over which diffusion needs to occur, the time required for sample molecules to interact with interior binding sites is also reduced. Diffusion is no longer limiting and flow rates can be dramatically increased - without compromising resolution or capacity. Separations can be achieved at 1,000 to 5,000 cm⁄hr compared to 50 to 360 cm⁄hr for conventional media.

POROS™ Ethyl RoboColumn (Thermo Scientific™)

This RoboColumn product is pre-packed with POROS Ethyl Hydrophobic Interaction Chromatography (HIC) Resin and suitable for impurity removal during large-scale downstream purification of biomolecules, primarily for bind/elute operations.

RoboColumn products are small chromatography columns that are provided in 8-column strips. They are useful for fully automated and parallel chromatographic separations using robotic liquid handling platforms.Features of POROS Ethyl HIC Resin include:

• Superior resolution capability while maintaining excellent pressure-flow properties, achieved by combining the high-flow characteristics of a POROS base bead with a 50-micron particle size. Higher flow rates help improve productivity.
• Unique selectivity due to the novel hydrophobic ligand, allowing for purification of a wide range of biomolecules, including therapeutic proteins, antibody fragments, antibody drug conjugates, and enzymes
• High capacity, reducing the process volume significantly for subsequent steps
• A rigid polymeric bead with covalent surface chemistry for easier handling and packing with superior physical and chemical stability, allowing cleaning with various clean-in-place (CIP) solutions such as sodium hydroxide and enabling a more robust downstream purification process

These features of POROS Ethyl HIC Resin are a major improvement to the performance of traditional HIC resins, setting the standard for hydrophobic interaction chromatography.

RoboColumn is a registered trademark of ATOLL GmbH.

Pierce™ Avidin Agarose (Thermo Scientific™)

Thermo Scientific Pierce Avidin Agarose can be used in a variety of applications for the affinity purification of biotinylated macromolecules.

Features of Avidin Agarose:

• Strong , nearly irreversible biotin:avidin interaction
• Binding biotinylated anti-transferrinfrom serum (1)
• Binding biotinyl peptides and elution with a SDS/Urea solution (2)
• Hybridization of biotinylated RNA to its complementary DNA and binding to immobilized avidin, with subsequent elution of the single-stranded DNA (3)
• Purification of double-stranded DNA (4)
• Approximate Binding Capacity >20 µg biotin/mL resin

In one variation, an antibody that has an affinity for a particular antigen is labeled with biotin. Cells containing the antigen are lysed, then incubated with the biotinylated antibody to form a typical antigen/antibody complex. To isolate the antigen, the crude mixture is passed through an immobilized avidin or streptavidin column, which will bind the complex. After appropriate washes, the antigen can then be eluted from the column with a pH 2.8 elution buffer. The biotinylated antibody is retained by the column.

Related Products
Pierce™ Monomeric Avidin Agarose

POROS™ CaptureSelect™ hCG Affinity HPLC Column, 2.1 x 30 mm (Thermo Scientific™)

POROS CaptureSelect hCG Affinity HPLC columns incorporate the advantages of high-speed POROS chromatography resins with the unique selectivity of CaptureSelect affinity ligands. CaptureSelect hCG Affinity HPLC columns recognize the alpha chain of human chorionic gonadotropin (hCG), enabling quantitation and small scale purification of hCG from the (recombinant) source material. Because it recognizes the alpha chain, it also binds LH, FSH, and TSH.

Analytical groups supporting biologic drug development require rapid, sensitive, and precise methods for protein quantitation from cell culture, fermentation harvests, or purification intermediates. It is critical for biopharmaceutical manufacturers to be able to monitor bioreactor product titers to determine when to harvest or abort a reactor cycle.

Features of POROS CaptureSelect hCG Affinity HPLC columns include:
Fast—POROS resins enable quantitation or purification of products in minutes
Hands-off operation—HPLC method can be automated for a streamlined operation
Robust—designed to deliver high-performance purification over hundreds of chromatography cycles

High speed with high specificity
POROS high performance chromatography allows for the quantitation of product from clarified harvest or partially purified intermediates—typically in under three minutes. Moreover, only minimal sample preparation (0.2 µm filtration) is required, and the eluted product is highly purified and suitable for further analysis, including mass spectrometry.

Automation
POROS Affinity HPLC columns are for use with HPLC systems, which allows the automation of chromatography methods, data collection, and data analysis. In addition, the immobilized affinity ligand enables one method and buffer system to be used across a class of molecules, streamlining analytical approaches.

Multiple cycles
Proper sample preparation, adherence to column operating instructions, and the use of an optimized column cleaning regimen results in strong re-use performance. POROS Affinity HPLC columns can be cycled hundreds of times for cost-effective and reproducible quantitation or purification.

Main characteristics

Protein binding capacity: 2.7 mg/mL
Species specificity: human
Column material: PEEK polymer
Frit: 2 µm, titanium
POROS resin: cross-linked poly(styrene-divinylbenzene)
CaptureSelect ligands: single-domain monospecific antibody fragment produced in Saccharomyces cerevisiae

CaptureSelect™ IgG-CH1 Affinity Matrix (Thermo Scientific™)

CaptureSelect™ IgG-CH1 Affinity Matrix has been specifically designed for the purification of recombinant human Fab fragments and IgGs, recognizing all IgG subclasses (1, 2, 3 and 4) independent of the light chain subclass (Kappa/Lambda).

Features of this affinity matrix include:

• Platform approach for purification of IgGs and their Fab fragments
• No co-purification of overexpressed free lights chains due to heavy chain binding
• Excellent scalability
• Non-animal-derived

CaptureSelect™ IgG-CH1 Affinity Matrix purifies IgG, Fab, and Fab2 fragments directly from complex source materials in a single step with high purity and yield.

Free of Animal Components
Our CaptureSelect™ products are affinity ligands created by a proprietary technology based on Camelid-derived single domain antibody fragments. The ligand is a 14 kDa single domain fragment comprising the 3 CDRs that form the antigen binding domain, efficiently produced by the yeast Saccharomyces cerevisiae in a production process free of animal components.

Main Characteristics:

Matrix: agarose-based, aldehyde-activated
Average particle size: 65 ± 10 µm
Ligand: CaptureSelect™ IgG-CH1 affinity ligand
Ligand coupling method: aldehyde coupling via NH2 residues of the ligand
Binding capacity: 25–30 g IgG/L resin depending on flow rate, column height and contact time
Elution conditions: 20 mM citric acid, 150 mM NaCl, pH 3.5
Flow characteristics: 200–400 cm/h
Formulation buffer: 20%(v/v) ethanol

HisPur™ Ni-NTA Superflow Agarose (Thermo Scientific™)

Thermo Scientific HisPur Ni-NTA Superflow Agarose is a nitrilotriacetic acid (NTA) modified Superflow 6 support charged with divalent nickel (Ni+2) designed for FPLC purification of poly-histidine-tagged proteins.

Features of HisPur Ni-NTA Superflow Agarose:

High capacity—binds greater than 20 mg of His-tagged GFP per mL of resin at a linear flow rate of 300 cm/hour
High purity—provides greater than 80% purity of eluted fractions when used to purify from lysates
Versatile—can be used to purify proteins under native and denaturing conditions
Robust—highly crosslinked beads tolerate linear flow rates up to 1200 cm/hour
Compatible—maintains function in a wide variety of chemicals and pH conditions
Cost effective—competitively priced resin is stable through multiple cycles of cleaning and reuse

HisPur Ni-NTA Superflow Agarose is a highly crosslinked, durable resin that does not compress at flow rates used for medium- or large-scale FPLC purifications. The resin holds up well to a variety of chemicals and pH values, and is compatible with common clean-in-place procedures. Compared to cobalt and other ligands used for immobilized metal affinity chromatography (IMAC), nickel provides greater capacity for His-tagged protein purification. HisPur Ni-NTA Superflow Agarose exhibits a high dynamic binding capacity across a range of flow rates, making it an excellent choice for larger scale purifications.

Applications:
• Medium- to large-scale FPLC purification of polyhistidine tagged proteins

Affinity chromatography is often used as a quick and easy approach for the purification of recombinant proteins. One such method of purification involves the binding of a poly-histidine tag to a divalent metal cation that has been coordinated by a chelator immobilized on a beaded support. For immobilized metal affinity chromatography (IMAC) purification of His-tagged proteins, the type of bead, chelator, and metal immobilized influences purity, yield, and flow rate performance of these purifications.

HisPur Ni-NTA Superflow Agarose is a nitrilotriacetic acid (NTA) modified Superflow 6 support charged with divalent nickel (Ni+2) designed for FPLC purification of poly-histidine-tagged protein. To demonstrate performance, 6xHis-GFP was over expressed in a 100L reactor and the cell mass was collected in multiple fractions. A portion of the biomass (240g) was lysed with 2L of lysis buffer supplemented with Thermo Scientific Halt Protease Inhibitor (Part No. 78439). The lysate was clarified and split into two fractions. The target protein was then purified using HisPur Ni-NTA Superflow Agarose and a leading competitor's resin in equivalent 200 mL packed bead columns. Total yield, recovery, and purity (>80%) were nearly identical for both resins, and more than 4 grams of target protein was purified in less than 3 hours.

More Product Data
Performance characterization of HisPur Ni-NTA Superflow Agarose

Related Products
HisPur™ Ni-NTA Resin