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 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

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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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 

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IEX 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. 

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Introduction

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

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

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 F Prepacked Chromatography Columns for Ion Exchange

PRC Prepacked Columns

Description

Part Number

Q Ceramic HyperD F 5x50, 1 mL, 1/pkg

PRC05X050QCHDF01

CM Ceramic HyperD F 5x50, 1 mL, 1/pkg

PRC05X050CMCHDF

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Q, S, DEAE, CM Ceramic HyperD® F - Ion Exchange Resins

PDF | 1.1 MB

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. 

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Introduction

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

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

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 F Prepacked Chromatography Columns for Ion Exchange

PRC Prepacked Columns

Description

Part Number

Q Ceramic HyperD F 5x50, 1 mL, 1/pkg

PRC05X050QCHDF01

CM Ceramic HyperD F 5x50, 1 mL, 1/pkg

PRC05X050CMCHDF01

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

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Q, S, DEAE, CM Ceramic HyperD® F - Ion Exchange Resins

PDF | 1.1 MB

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. 

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Introduction

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

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

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 F Prepacked Chromatography Columns for Ion Exchange

PRC Prepacked Columns

Description

Part Number

Q Ceramic HyperD F 5x50, 1 mL, 1/pkg

PRC05X050QCHDF01

CM Ceramic HyperD F 5x50, 1 mL, 1/pkg

PRC05X050CMCHDF01

AcroSep Prepacked Columns

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

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Q, S, DEAE, CM Ceramic HyperD® F - Ion Exchange Resins

PDF | 1.1 MB

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. 

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Salt Tolerant Advanced Recovery Anion Exchange Chromatography Sorbent

HyperCel STAR AX sorbent is an industry-scalable anion exchange chromatography sorbent 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 sorbent 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 sorbent is manufactured at large scale and meets the needs of industrial users and regulatory authorities. This sorbent 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 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.

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.

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 5x50 HyperCel STAR AX, 1 mL

PRCSTARAX1ML

PRC Column 8x100 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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HyperCel™ STAR AX Ion Exchange Resin Salt Tolerant Advanced Recovery Anion Exchange Chromatography Resin

PDF | 1.1 MB
IEX 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. 

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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. 

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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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Sartobind® Membrane Chromatography for Virus Purification

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ion exchange chromatography molecules