CIM® r-Protein G-8f mL Tube Monolithic Column
306KBAbout CIM® Protein G-8f Tube Monolithic Columns
The CIM® Protein G-8f Tube Monolithic Column consists of a unique polymeric monolith encased in a specially designed housing. These affinity columns are used for fast, highly efficient purification of immunoglobulin G. Protein G binds the Fc portion of antibodies (IgG class) without affecting antigen binding. In comparison to protein A, the range of polyclonal IgGs that bind strongly to protein G is much wider and adsorption takes place over a wide pH range. The design of these columns 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. This short separation layer length and the specially engineered highly porous structure allow operations at elevated flow rates with low-pressure drops (only a few bar).
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.
| Catalogue number: | 437.1011 |
| Tube chemistry: | affinity, protein G from Streptococcus aureus |
| 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 1.0 mg/mL, 20 mM Tris-HCl buffer, pH 7.4, flow rate 8 mL/min) |
| Working flow rates: | up to 200 mL/min (uav = 671 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® Protein G-8f Tube Monolithic Columns
Regeneration
To regenerate (to wash out from the monolithic column any strong or unspecifically bound substances), wash the column with at least 10 column volumes of a 0.1 M buffer containing 1 M NaCl, pH 7.0–8.0 at one-half of the working flow rate. After that, wash the column with at least 10 column volumes of a 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 a complete blockage of the column. A specific CIP protocol should be designed according to the type of contaminants that are present in your sample. However, in most cases, the followingprocedures 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 to 0.5 M buffer) at the 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 or 70 % ethanol 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® Protein G-8f Tube Monolithic Columns
CIM&® Protein G-8f Tube Monolithic Column be stored at a temperature of +4°C to +8°C in the presence of a suitable bacteriostatic agent, e. g. 20% ethanol. |
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