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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

Resins
Attributes Ceramic HyperD Q Ceramic HyperD DEAE Ceramic HyperD CM 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)
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Membranes
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 
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Ion Exchange Resins

Ceramic HyperD Q

Ceramic HyperD Q 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. 

Description

Q, DEAE and CM Ceramic HyperD™ F ion exchange chromatography  sorbents 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

Applications

  • Recombinant proteins
  • Plasmid purification
  • Purification of enzymes, plasma proteins
  • Vaccines
  • Direct capture of monoclonal antibodies (CM Ceramic HyperD™ F)
  • IgM

Specifications

Item Q Sorbent DEAE Sorbent CM Sorbent
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

Ordering Information

Ceramic HyperD F Sorbents
Q Sorbent DEAE Sorbent CM Sorbent
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
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Ceramic HyperD DEAE

Ceramic HyperD DEAE 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. 

 

Description

Q, DEAE and CM Ceramic HyperD™ F ion exchange chromatography  sorbents 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

Applications

  • Recombinant proteins
  • Plasmid purification
  • Purification of enzymes, plasma proteins
  • Vaccines
  • Direct capture of monoclonal antibodies (CM Ceramic HyperD™ F)
  • IgM

Specifications

Item Q Sorbent DEAE Sorbent CM Sorbent
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

Ordering Information

Ceramic HyperD F Sorbents
Q Sorbent DEAE Sorbent CM Sorbent
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
Request a Quote

Ceramic HyperD CM

Ceramic HyperD CM 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. 

 

Description

Q, DEAE and CM Ceramic HyperD™ F ion exchange chromatography  sorbents 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

Applications

  • Recombinant proteins
  • Plasmid purification
  • Purification of enzymes, plasma proteins
  • Vaccines
  • Direct capture of monoclonal antibodies (CM Ceramic HyperD™ F)
  • IgM

Specifications

Item Q Sorbent DEAE Sorbent CM Sorbent
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

Ordering Information

Ceramic HyperD F Sorbents
Q Sorbent DEAE Sorbent CM Sorbent
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
Request a Quote

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. 

 

Description

Applications

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 performed on S HyperCel resin, 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 (S HyperCel resin).

This last polishing step on S HyperCel resin led to a purified fraction of HSA eluted at pH 7.0 with purity >99%, and a capacity around 65 mg/mL.

Load Capacity (mg/mL) Yield Purity
Capture on HyperCel STAR AX resin Undiluted plasma > 30 mg/mL 90% 99%
Polishing on S HyperCel resin pH 4.0 eluate from HyperCel STAR AX resin 65 mg/mL 95% 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 resinbefore 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.

Specifications

HyperCel STAR AX sorbent 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 sorbent 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 sorbent 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 sorbent facilitates the sorbent transfer, avoiding the agitation and suspension of large material volumes).

HyperCel STAR AX sorbent 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.

Ordering Information

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
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HyperCel STAR AX PRC Prepacked Columns

Description Part Number
PRC Column 5×50 HyperCel STAR AX, 1 mL PRCSTARAX1ML
PRC Column 8×100 HyperCel STAR AX, 5 mL PRCSTARAX5ML
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Description Part Number
AcroPrep ScreenExpert Plates filled with HyperCel STAR AX Sorbent 96WPSTARAX50
ScreenExpert RoboColumn HyperCel STAR AX 200 μL, row of 8 SR2STARAX
96-well RoboColumn array plate SR96WAP
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Ion Exchange Membranes

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. 

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