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GE17-5080-01

HiTrap® rprotein A Fast Flow

GE Healthcare, 17-5080-01, pack of 1 × 5 mL

  •  NACRES NA.56

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Properties

Related Categories Affinity Chromatography, Analytical/Chromatography, GE Healthcare Life Sciences Protein Sample Preparation Formats, Heparin, HiTrap & PD-10 Cartridges,
shelf life   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.
packaging   pack of 1 × 5 mL
mfr. no.   GE Healthcare, 17-5080-01
parameter   <20 mL/min flow rate (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.)
  42 psi
bed size   16 mm × 25 mm
bed volume   5 mL
column I.D.   16 mm
matrix   4% cross-linked agarose
particle size   60-165 μm
average diameter   90 μm (d50v)
cleaning   2 - 11 (pH below 3 is sometimes required to elute strongly bound IgG species. However, protein ligands may hydrolyze at very low pH.)
working range   3 - 10
capacity   ~50 mg binding capacity (human lgG/ml medium)(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.)
suitability   suitable for bioprocess medium
storage temp.   2-8°C

Description

General description

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.

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.

Analysis Note

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

Legal Information

HiTrap is a registered trademark of Cytiva

Sepharose is a registered trademark of Cytiva

Safety & Documentation

Safety Information

RIDADR 
UN1170 - class 3 - PG 3 - Ethanol, solution

Documents

Certificate of Analysis (COA)

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Protocols & Articles

Articles

Performing a Purification of IgG Antibodies with HiTrap® Protein G HP Columns

HiTrap® Protein G HP (Figure 3.10) is a convenient, ready-to-use column prepacked with Protein G Sepharose® High Performance. The columns are available in 1 ml and 5 ml sizes. In common with all HiTr...
Keywords: Affinity chromatography, Buffers, Cell culture, Chromatography, Immunodiffusion, Ligands, PAGE, Sample preparations

Performing a Purification of IgG Antibodies with HiTrap® rProtein A FF columns

HiTrap rProtein A FF is a ready-to-use column prepacked with rProtein A Sepharose Fast Flow (Figure 3.20) for the convenient purification of monoclonal antibodies from cell culture supernatants, serum...
Keywords: Affinity chromatography, Cell culture, Chromatography, PAGE, Sample preparations

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

HiTrap™ IgM Purification HP 1 ml columns are prepacked with a thiophilic adsorption medium (2-mercaptopyridine coupled to Sepharose® High Performance). The binding capacity of HiTrap™ IgM Purification ...
Keywords: Adsorption, Affinity chromatography, Cell culture, Chromatography, Enzyme-linked immunosorbent assay, Ligands, PAGE, Precipitation, Purification, Sample preparations

Purification using protein A chromatography media

Protein A is derived from a strain of Staphylococcus aureus and contains five regions that bind to the Fc region of IgG. As an affinity ligand, protein A is coupled to Sepharose® so that these region...
Keywords: Affinity chromatography, Buffers, Chromatography, Fermentation, Immunoprecipitation, Ligands, Purification, Size-exclusion chromatography

Selection Guide for Polymeric Resins in Chromatography & Purification

Sigma-Aldrich carries a vast array of resins, from leading manufacturers; such as, Dow Chemical, Rohm & Haas, Mitsubishi, and Lanxess. Our product line includes adsorbent, ion-exchange (anion / catio...
Keywords: Adsorption, Antibiotics, Combinatorial synthesis, Detergents, Fermentation, Filtration, Hormones, Ion Exchange, Ion exchange resins, Pharmaceutical, Purification, Separation

Protocols

Affinity Chromatography Troubleshooting

This section focuses on practical problems that may occur when running a chromatography column. The diagrams below give an indication of how a chromatogram may deviate from the ideal during affnity p...
Keywords: Adsorption, Affinity chromatography, Buffers, Chromatography, Detergents, Digestions, Fractionation, Ligands, PAGE, Precipitation, Purification, Sample preparations, Separation, Solvents

Buffer Exchange and Desalting for Affinity Chromatography

Dialysis is frequently mentioned in the literature as a technique to remove salt or other small molecules and to exchange the buffer composition of a sample. However, dialysis is generally a very slo...
Keywords: Absorption, Affinity chromatography, Buffers, Chromatography, Dialysis, Separation

Column Packing and Preparation for Affinity Chromatography

Appendix 3,  Extracted from Affinity Chromatography Principles and Methods, GE Healthcare, 2007
Keywords: Affinity chromatography, Antimicrobials, Buffers, Chromatography, Separation

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

Appendix 4 extracted from Affinity Chromatography Principles and Methods, GE Healthcare, 2007
Keywords: Affinity chromatography, Chromatography

Performing a Separation with GE Healthcare Products Based on Protein A

Protein A is derived from a strain of Staphylococcus aureus and contains five regions that bind to the Fc region of IgG. As an affinity ligand, protein A is coupled to Sepharose so that these regions a...
Keywords: Affinity chromatography, Buffers, Cell culture, Chromatography, Ligands, PAGE, Purification

Sample Preparation for Affinity Chromatography

Appendix 1 extracted from Affinity Chromatography Principles and Methods, GE Healthcare, 2007
Keywords: Adsorption, Affinity chromatography, Buffers, Centrifugation, Chromatography, Detergents, Filtration, Nucleic acid denaturation, PAGE, Precipitation, Sample preparations

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