Ion Exchange Chromatography
Ion Exchange Chromatography (IEX) is the technique used to separate molecules based on their electrical charge. Positively charged molecules will bind to a cation exchanger (CEX), and negatively charged molecules will bind to an anion exchanger (AEX).
The most common application for IEX is polishing of the target molecule, but it can also be used to capture the product such as viral vectors.
Find the Right Solution for Your Ion Exchange Chromatography Process
Ion exchange resins are applied for the removal of impurities and exhibit good binding capacities for small and medium-size proteins. The salt-tolerant Hypercel Star AX anion exchanger, in addition, offers simple process integration without the need for feedstream manipulation.
Attributes | Hypercel Star AX | |||
---|---|---|---|---|
Particle diameter | 50 µm | 50 µm | 50 µm | 80 µm |
Dynamic Binding Capacity (10 % BT) | > 85 mg/mL BSA | > 85 mg/mL BSA | > 60 mg/mL IgG | > 100 mg/mL BSA |
Functionality | Strong Anion Exchange | Weak Anion Exchange | Weak Cation Exchange | Weak Anion Exchange (Salt tolerant) |
Ion exchange membranes are also used for polishing of mAbs. Compared to resins, higher flow rates can be applied to membranes resulting in shorter process time and higher productivity. Further, membranes have a higher binding capacity to virus, virus-like particles, and other large complexes compared to resins. Therefore, they are widely used in capture of these molecules. Our membranes are provided ready-to-use devices.
Attributes | Sartobind® Q | Sartobind STIC® | Sartobind® S |
---|---|---|---|
Pore size | 3-5 µm | 3-5 µm | 3-5 µm |
Functionality | Strong Anion Exchange | Weak Anion Exchange (Salt tolerant) | Strong Cation Exchange |
Q Ceramic HyperD®
Q Ceramic HyperD® is a strong anion exchanger that is suited for removal of anionic impurities (i.e., host cell protein and DNA). Its 50 µm base bead offers a high resolution for polishing applications when required.
Introduction
Q, DEAE and CM Ceramic HyperD® F ion exchange chromatography resins are designed using proprietary 'gel-in-a-shell' design that provides rapid mass transfer and effective binding. It relies on a high capacity hydrogel, polymerized within the gigapores of a rigid ceramic bead.
Features and Benefits
- High dynamic binding capacity at high flow rates
- Truly rigid, non-compressible resin
- Salt tolerant CM Ceramic HyperD® F resin reduces Ultrafiltration/Diafiltration (UF/DF) requirements
- Recombinant proteins
- Plasmid purification
- Purification of enzymes, plasma proteins
- Vaccines
- Direct capture of monoclonal antibodies (CM Ceramic HyperD® F)
- IgM
Item | Q Resin | DEAE Resin | CM Resin |
Average particle size (µm) | 50 | 50 | 50 |
Dynamic Binding Capacity (mg/mL) 10% breakthrough at 200 cm/hr | BSA* > 85¹ | BSA* > 85¹ | IgG > 602 |
Amount of ionic groups (µEq/mL) | > 250 | > 200 | 250 - 400 |
Working pH | 2 -12 | ||
Cleaning pH | 1 - 14 | ||
Pressure resistance | 70 barg (1,000 psi) | ||
Volumes changes due to pH and ionic strength | Non compressible |
* BSA = Bovine Serum Albumin
¹ 5 mg/mL BSA in 50 mM Tris-HCl, pH 8.6
25 mg/mL hu IgG in 50 mM sodium acetate, 100 mM NaCl, pH 4.7
Ceramic HyperD® F Resins | |||
Pack Size | Part Number | ||
25 mL | 20066-031 | 20067-039 | 20050-035 |
100 mL | 20066-023 | 20067-021 | 20050-027 |
1 L | 20066-015 | 20067-013 | 20050-019 |
5 L | 20066-064 | 20067-054 | 20050-050 |
10 L | 20066-056 | 20067-047 | 20050-043 |
Ceramic HyperD® F Prepacked Chromatography Columns for Ion Exchange | |
PRC Prepacked Columns | |
Description | Part Number |
Q Ceramic HyperD F 5x50, 1 mL, 1/pkg | |
CM Ceramic HyperD F 5x50, 1 mL, 1/pkg |
DEAE Ceramic HyperD®
DEAE Ceramic HyperD® is a weak anion exchanger for polishing purification. Changes in conductivity and pH can optimize purification steps by utilizing the charge change on protein and ligand.
Introduction
Q, DEAE and CM Ceramic HyperD® F ion exchange chromatography resins are designed using proprietary 'gel-in-a-shell' design that provides rapid mass transfer and effective binding. It relies on a high capacity hydrogel, polymerized within the gigapores of a rigid ceramic bead.
Features and Benefits
- High dynamic binding capacity at high flow rates
- Truly rigid, non-compressible sorbent
- Salt tolerant CM Ceramic HyperD F sorbent reduces Ultrafiltration/Diafiltration (UF/DF) requirements
- Recombinant proteins
- Plasmid purification
- Purification of enzymes, plasma proteins
- Vaccines
- Direct capture of monoclonal antibodies (CM Ceramic HyperD® F)
- IgM
Item | Q Resin | DEAE Resin | CM Resin | |
Average particle size (µm) | 50 | 50 | 50 | |
Dynamic Binding Capacity (mg/mL) 10% breakthrough at 200 cm/hr | BSA* > 85¹ | BSA* > 85¹ | IgG > 602 | |
Amount of ionic groups (µEq/mL) | > 250 | > 200 | 250 - 400 | |
Working pH | 2 -12 | |||
Cleaning pH | 1 - 14 | |||
Pressure resistance | 70 barg (1,000 psi) | |||
Volumes changes due to pH and ionic strength | Non compressible |
* BSA = Bovine Serum Albumin
¹ 5 mg/mL BSA in 50 mM Tris-HCl, pH 8.6
25 mg/mL hu IgG in 50 mM sodium acetate, 100 mM NaCl, pH 4.7
Ceramic HyperD® F Resins | |||
Pack Size | Part Number | ||
25 mL | 20066-031 | 20067-039 | 20050-035 |
100 mL | 20066-023 | 20067-021 | 20050-027 |
1 L | 20066-015 | 20067-013 | 20050-019 |
5 L | 20066-064 | 20067-054 | 20050-050 |
10 L | 20066-056 | 20067-047 | 20050-043 |
Ceramic HyperD® F Prepacked Chromatography Columns for Ion Exchange | |
PRC Prepacked Columns | |
Description | Part Number |
Q Ceramic HyperD F 5x50, 1 mL, 1/pkg | |
CM Ceramic HyperD F 5x50, 1 mL, 1/pkg | |
AcroSep Prepacked Columns | |
Description | Part Number |
Q Ceramic HyperD F, 1 mL, Red, 5/pkg | 20066-C001 |
CM Ceramic HyperD F, 1 mL, Green, 5/pkg | 20050-C001 |
DEAE Ceramic HyperD F, 1 mL, Orange, 5/pkg | 20067-C001 |
(1) each: Q, CM, and DEAE Ceramic HyperD F, 1 mL, 4/pkg | IEXVP-C001 |
CM Ceramic HyperD®
CM Ceramic HyperD® is a weak cation exchanger for polishing applications. Changes in conductivity and pH can optimize purification steps by utilizing the charge change on protein and ligand.
Introduction
Q, DEAE and CM Ceramic HyperD® F ion exchange chromatography resins are designed using proprietary 'gel-in-a-shell' design that provides rapid mass transfer and effective binding. It relies on a high capacity hydrogel, polymerized within the gigapores of a rigid ceramic bead.
Features and Benefits
- High dynamic binding capacity at high flow rates
- Truly rigid, non-compressible resin
- Salt tolerant CM Ceramic HyperD® F resin reduces Ultrafiltration/Diafiltration (UF/DF) requirements
- Recombinant proteins
- Plasmid purification
- Purification of enzymes, plasma proteins
- Vaccines
- Direct capture of monoclonal antibodies (CM Ceramic HyperD® F)
- IgM
Item | Q Resin | DEAE Resin | CM Resin | |
Average particle size (µm) | 50 | 50 | 50 | |
Dynamic Binding Capacity (mg/mL) 10% breakthrough at 200 cm/hr | BSA* > 85¹ | BSA* > 85¹ | IgG > 602 | |
Amount of ionic groups (µEq/mL) | > 250 | > 200 | 250 - 400 | |
Working pH | 2 -12 | |||
Cleaning pH | 1 - 14 | |||
Pressure resistance | 70 barg (1,000 psi) | |||
Volumes changes due to pH and ionic strength | Non compressible | |||
* BSA = Bovine Serum Albumin
¹ 5 mg/mL BSA in 50 mM Tris-HCl, pH 8.6
25 mg/mL hu IgG in 50 mM sodium acetate, 100 mM NaCl, pH 4.7
Ceramic HyperD® F Resins | |||
Pack Size | Part Number | ||
25 mL | 20066-031 | 20067-039 | 20050-035 |
100 mL | 20066-023 | 20067-021 | 20050-027 |
1 L | 20066-015 | 20067-013 | 20050-019 |
5 L | 20066-064 | 20067-054 | 20050-050 |
10 L | 20066-056 | 20067-047 | 20050-043 |
Ceramic HyperD® F Prepacked Chromatography Columns for Ion Exchange | |
PRC Prepacked Columns | |
Description | Part Number |
Q Ceramic HyperD F 5x50, 1 mL, 1/pkg | |
CM Ceramic HyperD F 5x50, 1 mL, 1/pkg | |
Q Ceramic HyperD F, 1 mL, Red, 5/pkg | 20066-C001 |
CM Ceramic HyperD F, 1 mL, Green, 5/pkg | 20050-C001 |
DEAE Ceramic HyperD F, 1 mL, Orange, 5/pkg | 20067-C001 |
(1) each: Q, CM, and DEAE Ceramic HyperD F, 1 mL, 4/pkg | IEXVP-C001 |
Hypercel Star AX
Hypercel Star AX is a salt-tolerant anion exchanger with a primary amine as functional group. It can easily be integrated into a purification workflow without feed stream manipulation.
Salt Tolerant Advanced Recovery Anion Exchange Chromatography Resin
HyperCel STAR AX resin is an industry-scalable anion exchange chromatography resin designed for high productivity protein capture at moderate or higher salt conductivity (2 to 15 mS/cm), typical of undiluted biological feedstocks (i.e., mammalian cell culture supernatants, E. coli feedstock, plasma, others…).
Applications include direct capture of recombinant proteins, monoclonal and polyclonal antibodies, plasma derivatives or other biopharmaceuticals.
Due to its ability to capture directly proteins from undiluted feedstreams, HyperCel STAR AX resin can also be used for early contaminant removal (i.e., CHO Host Cell Proteins), before target purification, i.e., before MAb capture by a Protein A affinity step.
HyperCel STAR AX resin is manufactured at large scale and meets the needs of industrial users and regulatory authorities. This resin offers:
- High dynamic binding capacity (DBC) at short residence time (2 minutes or lower)
- Direct capture of protein from undiluted feedstock at moderate or high conductivity
- Excellent flow rate properties for fast feedstock processing
- Distinctive selectivity, consistent in a broad conductivity range (2 – 15 mS/cm)
- Enhanced process economics
Application 1. Direct Capture of Albumin from Undiluted Plasma
The objective is to evaluate HyperCel STAR AX resin as the first step in a two-step purification sequence to capture human serum albumin (HSA) from undiluted plasma (conductivity 11 mS/cm). HyperCel STAR AX resin was used as a first capture step, followed by an orthogonal cation exchange step, without pH or conductivity adjustment of the plasma. Neat undiluted plasma (pH 7.6, 11mS/cm), was loaded with DBC of 30 mg/mL.
Design of Experiments (DoE) in 96-well filter plates was performed to determine optimal conditions to achieve the best yield/purity ratio.
These conditions were then transferred to column chromatography on PRC prepacked columns.
Conditions determined in 96-well plates were applied to chromatography of undiluted plasma on HyperCel STAR AX resin in a 1 mL PRC prepacked column. Chromatogram and SDS-PAGE analysis of fractions confirmed that most contaminants were eliminated by high conductivity wash, leading to a 99% pure HSA fraction in a single step, with 90% yield.
In addition, HyperCel STAR AX resin used at capture step with undiluted plasma had a DBC >30 mg/mL, more than 2-fold higher than that of a conventional DEAE agarose resin tested in these conditions.
HSA was eluted by a simple decrease of pH (4.0), without addition of salt, allowing a direct load on an orthogonal cation exchange column.
Load | Capacity (mg/mL) | Yield | Purity | |
Capture on HyperCel STAR AX resin | Undiluted plasma | > 30 mg/mL | 90% | 99% |
Application 2. Early Removal of CHOPs before Protein A Capture of a Monoclonal Antibody (MAb)
The objective of this study was to evaluate the impact of a pre-purification step using HyperCel STAR AX resin before conventional Protein A resin capture of a monoclonal antibody (MAb) from mammalian cell culture feedstock.
The content of contaminating CHOPs (Chinese Hamster Ovary Proteins) was compared using a commercial ELISA assay for the two purification schemes.
Data shows that using HyperCel STAR AX resin prior to Protein A results in a better CHOP reduction (>8-fold). Due to this synergistic effect, starting from an initial Host Cell Protein (HCP) content of 413,000 ppm, the final CHOP level is reduced to ∼400 ppm (3-Log reduction), with a MAb recovery of 90%.
A pre-purification step can impact positively process economics by extending the lifetime of an expensive Protein A column used as standard step for MAb capture.
HyperCel STAR AX resin is composed of a rigid cellulose matrix that has excellent flow properties and generates low backpressure, compatible with the needs of manufacturing-scale protein production. The resin is available in a variety of packaging configurations as well as convenient 1 mL and 5 mL PRC prepacked columns designed for fast method optimization, selectivity screening or small preparative work. HyperCel STAR AX resin is supplied as a slurry/suspension in 1 M NaCl containing 20% (v/v) ethanol or as a moist cake for process-scale applications (the moist cake resin facilitates the resin transfer, avoiding the agitation and suspension of large material volumes).
HyperCel STAR AX resin has a chemical stability that ensures simple clean-in-place (CIP) and storage. For standard CIP, 0.5 to 1 M NaOH treatment is recommended, while long-term storage in 10 to 100 mM NaOH is possible.
Main Properties
Average particle size | 80 µm |
Ion exchange ligand | Primary amine |
Dynamic binding capacity1 | >100 mg BSA/mL within pH range 7.5 – 8.0 and conductivity 15 mS/cm |
Typical operating range of feedstock conductivity | 2 – 15 mS/cm |
Recommended cleaning conditions2 | 1 M NaOH |
1 Determined using a 5 mg/mL BSA in 25 mM Tris-HCl , 0.14 M NaCl at 2 minute residence time.
2 Injection of 5 column volumes (CV) of 0.5 – 1 M NaOH, 1 hour contact time.
HyperCel STAR AX Resin
Size | Part Number |
25 mL | 20197-026 |
100 mL | 20197-032 |
1 L | 20197-046 |
5 L | 20197-058 |
10 L | 20197-064 |
HyperCel STAR AX PRC Prepacked Columns
Description | Part Number |
PRC Column 5x50 HyperCel STAR AX, 1 mL | PRCSTARAX1ML |
PRC Column 8x100 HyperCel STAR AX, 5 mL | PRCSTARAX5ML |
Description | Part Number |
RoboColumn HyperCel STAR AX 200 μL, row of 8 | SR2STARAX |
RoboColumn HyperCel STAR AX 600 µL, row of 8 | SR6STARAX |
Sartobind® Q
Sartobind® Q is a strong basic anion exchanger with quaternary ammonium as ligand. This membrane is provided in ready-to-use devices with two different bed heights.
Sartobind STIC®
Sartobind STIC® PA is a salt-tolerant weak basic anion exchanger. The ligand is a primary amine. Due to its salt-tolerance, it efficiently works at higher conductivity. This membrane is provided in ready-to-use devices in standard bed height.
Sartobind® S
Sartobind® S is a strong acidic cation exchanger with a sulfonic acid as ligand. This membrane is provided in ready-to-use devices with two different bed heights.