CIM® r-Protein A-8f mL Tube Monolithic Column
About CIM® r-Protein A-8f Tube Monolithic Column
The CIM® r-Protein A-8f high throughput Tube Monolithic Column consists of a unique polymeric monolith encased in a specially designed housing. This column is used for extremely fast, highly efficient purification of immunoglobulin G. Protein A binds to Fc portion of antibodies (IgG class) without affecting antigen binding. The design of the column allows the mobile phase to run in a radial direction from the outer to the inner surface. In this way, resolution is preserved during scale-up while maintaining the short separation layer length. The short monolithic length and the specially engineered highly porous structure allow operations at elevated flow rates with low-pressure drops (only few bars).
The following information is being provided to ensure proper product care and maximal product life. All other information can be found in the Instruction Manual that accompanied the product.
| Catalog number: | 437.1004 |
| Tube chemistry: | affinity, protein A from Staphylococcus aureus |
| Ligand chemistry: | 0.22 ± 0.02 mmol/g dry support |
| Support matrix: | poly (glycidyl methacrylate-co-ethylene dimethacrylate) |
| Tube dimensions: | outer diameter: 15.0 mm; inner diameter: 6.5 mm; length: 56.0 mm; bed volume: 8.0 mL |
| Dynamic binding capacity: | ≥ 9 mg/mL Conditions: human IgG 1 mg/mL, 20 mM Tris-HCl buffer, pH 7.4, flow rate 8 mL/min |
| Working flow rates: | up to 400 mL/min (uav = 1340 cm/h) |
| Working system pressure: | up to 20 bar (2 MPa); WARNING: Do not exceed the maximum allowed pressure as this might seriously damage your column! |
| Temperature stability: | 4°C (39°F) to 40°C (104°F) WARNING: Avoid prolonged use at elevated temperatures! |
| Recommended pH: | working range 2–11 cleaning-in-place 2–13 |
Caring for the CIM® r-Protein A-8f Tube Monolithic Column
Regeneration
To regenerate (i.e. to wash out any strong or unspecifically bound substances from the monolithic column), wash the column with at least 10 column volumes of a buffer containing 1 M NaCl, pH 7-8, at one-half of the working flow rate. Then, wash the column with at least 10 column volumes of low pH solution (e.g. 10 mM HCl or 0.1 M glycine-HCl) at one half of the working flow rate. Re-equilibrate the column with at least 10 column volumes of the working mobile phase at the working flow rate. For best results, this should be done at the end of each chromatographic run.Cleaning In Place (CIP)
In some cases, the simple regeneration of the monolithic column is not enough. Sample molecules may not fully elute from the column or may even precipitate on the column. This build up of contaminants on the monolithic column may cause loss of resolution and binding capacity, increased back pressure, or complete blockage of the column. A specific CIP protocol should be designed according to the type of contaminants that are present in the sample. However, in most cases, the following procedures can be used:
1. Removal of precipitated proteins
- Wash with 5–10 column volumes of a 0.1 M NaOH.
Use low enough flow rates to expose the column to the cleaning reagent for several minutes. - Wash with 10 column volumes of deionized water at the working flow rate.
- Wash with 10 column volumes of a concentrated buffer (e.g. 0.1 - 0.5 M buffer) at working flow rate in order to restore the appropriate pH.
- Re-equilibrate with 10 column volumes of the working mobile phase (buffer) at the working flow rate.
- Wash with 5–10 column volumes of deionized water at one-half of the working flow rate.
- Wash with 5–10 column volumes of a 30 % 2-propanol at one-half of the working flow rate.
- Wash with 5 column volumes of deionized water at the working flow rate.
- Re-equilibrate with 10 column volumes of the working mobile phase (buffer) at the working flow rate.
Storage of CIM® r-Protein A-8f Tube Monolithic Columns
CIM® r-Protein A-8f Tube Monolithic Column should be stored at +4 °C (39 °F) to +8 °C (46 °F) in the presence of a suitable bacteriostatic agent, e.g. 20 % ethanol. |
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