Sepapure protein A FPLC column
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Sepapure protein A FPLC column Short guide

Note:  For your own safety, read the instructions and observe the warnings and safety information on the device and in the instructions. Keep the instructions for future reference. KNAUER Technical Support: Version information: Phone: +49 30 809727-111 (9-17h, Central European Time) Fax: +49 30 8015010 E-Mail: support@knauer.net Languages: German, English KNAUER Wissenschaftliche Geräte GmbH Hegauer Weg 38 14163 Berlin Germany Phone: +49 30 809727-0 Fax: +49 30 8015010 Internet: www.knauer.net E-Mail: info@knauer.net Article number: V6011 Version number: 1.0 Last update: 2019/01/29 The...

Table of Contents 1. Specifications. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 1.1 Sepapure FPLC columns short guide, V6011

Specifications Definition Sepapure Protein A columns are designed for the purification of antibodies and antibody fragments via the Protein A – Fc region interaction. The columns are designed to be used with low-pressure FPLC-type automated purification systems and operated below 3 bar (0.3 MPa, 2.96 atm). Hardware specifications Column housing Polyethylene (nominal 20 µm porosity) Resin specifications Resin name Base matrix material 4% cross-linked, beaded agarose Mean bead diameter Ligand density Static antibody binding capacity Flow rate pH stability (with metal ion loaded) 2 – 10 (short...

Sepapure columns are designed to be used with most aqueous phase chromatography buffers. A suggested buffer system is shown below, although other buffers may be used. Please check the resin specifications for further details. Equilibration buffer Wash buffer Elution buffer 0.1 M glycine, pH 3.0 or 0.1 M citric acid, pH 3.0 2. Preparing the column Sepapure Protein A columns are supplied with 20% ethanol as the storage buffer. This must be removed prior to purification. Process Remove the end-plugs and connect the column to the control system, taking care to avoid introduction of air into the...

3 Elute the protein using the selected elution buffer. A stepwise or linear gradient may be used to determine the precise elution point of the target antibody. Antibodies can be eluted in a pH range starting from 5.5 and decreasing to as low as 3.0. Strongly bound antibodies may require an elution pH of between 2.0 and 3.0. Initially, all elution fractions should be collected for further analysis. Buffer exchange and/or desalting might be required following elution and we recommend Sepapure Desalting Columns (010X460SPZ and 020X460SPZ) for this purpose. Acid-labile antibodies should be...

Latest KNAUER instructions online: https://www.knauer.net/en/Support/User-manuals KNAUER Wissenschaftliche Geräte GmbH Hegauer Weg 38 14163 Berlin Phone: +49 30 809727-0 Fax: +49 30 8015010 E-Mail: info@knauer.net Internet: www.knauer.net