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Merck
CN

GE17-5080-02

HiTrap® rProtein A Fast Flow

Cytiva 17-5080-02, pack of 5 × 5 mL

Synonym(s):

affinity column, chromatography column, mAb purification column, prepacked column, protein A column

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About This Item

NACRES:
NA.56
UNSPSC Code:
23151816

Key Documents

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Technical Service
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storage condition

(20% Ethanol)

separation technique

affinity

storage temp.

2-8°C

form

prepacked column

crosslinking

cross-linked

packaging

pack of 5 × 5 mL

manufacturer/tradename

Cytiva 17-5080-02

parameter

0.5 Mpa max. pressure, 20 mL/min max. flow rate, 5 mL/min flow rate

technique(s)

liquid chromatography (LC): suitable

bed volume

5 mL

column I.D.

16 mm × 25 mm

matrix

4% cross-linked agarose

matrix active group

recombinant protein A ligand (E. coli)

particle size

~90 μm

cleaning

3-12

operating pH

3-10

capacity

~35 mg binding capacity (human lgG / ml medium)

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General description

Capacity of HiTrap rProtein A for some monoclonal antibodies. Running conditions: Binding buffer: 20 mM sodium phosphate (incl. 3 M NaCl for IgG), pH 7.0. Elution buffer: 0.1 M sodium citrate, pH 3.0.
HiTrap® rProtein A FF columns are prepacked with rProtein A Sepharose™ Fast Flow for purification and fractionation of IgG subclasses and IgG from different sources, including cell supernatants and ascites.

The recombinant protein A ligand (produced in E. coli) is coupled to Sepharose™ Fast Flow by a technique which generates a stable thioether linkage between rProtein A and the base matrix. The coupling technique is optimized to give high binding capacity for IgG.
pH below 3 is sometimes required to elute strongly bound IgG species. However, protein ligands may hydrolyze at very low pH.

Features and Benefits

  • Quick, convenient preparative purification of monoclonal antibodies.
  • Well-established protein A-based medium used in many approved MAb processes.
  • Recombinant protein A exhibits similar Fc region specificity to that of native protein A but shows enhanced binding capacity and fewer regulatory concerns due to the total absence of mammalian culture in the ligand production and purification.
  • Simple operations with a syringe, pump, or high-performance chromatography system, such as AKTA design.

Preparation Note

Please be aware this product may be shipped 90 days before the expiration date. For more information on the batch specific expiration date, please contact technical service.

Analysis Note

To view the Certificate of Analysis for this product, please visit www.cytiva.com.

Other Notes

The pressure over the packed bed varies depending on a range of parameters such as the characteristics of the chromatography medium and the column tubing used.

Legal Information

Sepharose is a trademark of Cytiva
HiTrap is a registered trademark of Cytiva

pictograms

Flame
GHS02

signalword

Warning

hcodes

H226

Storage Class

3 - Flammable liquids

Regulatory Information

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Articles

Affinity Chromatography Troubleshooting

This page shows how to solve practical problems that may occur when running an affinity chromatography column.

Performing a Purification of IgM Antibodies with HiTrap™ IgM Purification HP

This page shows how to purify IgM antibodies by affinity chromatography using HiTrap™ IgM Purification HP.

Purification Using Protein A-based Chromatography Media

This page shows various purification options for Protein A Sepharose chromatography media and describes typical binding and elution conditions for Protein A Sepharose chromatography media.

Protocols

Protein A-Based Separation with Cytiva Products

Protein A, coupled to Sepharose, binds IgG via its Fc regions, facilitating affinity chromatography purification.

Buffer Exchange and Desalting for Affinity Chromatography

How to perform buffer exchange and desalting with Sephadex G-25, HiTrap Desalting columns, or ÄKTAprime plus.

Column Packing and Preparation Protocol for Affinity Chromatography

This page shows how to perform column packing and preparation for affinity chromatography when using Tricorn™ or XK columns available from Cytiva.

Converting From Linear Flow (cm/hour) to Volumetric Flow Rates (mL/min) and Vice Versa

This page shows how to convert between linear flow and volumetric flow rates in affinity chromatography.

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