CIM® r-Protein L Disk Monolithic Column
396KBAbout the CIM® r-Protein L Disk
The CIM® r-Protein L Disk consists of a unique monolithic stationary phase that is easily placed in a dedicated plastic housing forming a CIM® Disk Monolithic Column. This affinity column is used for fast, highly efficient purification of species containing certain types of kappa light chains. Protein L binds all classes of Ig (IgG, IgM, IgA, IgE and IgD) and also fragments containing kappa light chains regardless of heavy chain subclassa,b. 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.
| Color code: | red with one line |
| Catalog number: | 217.1021 |
| Disk chemistry: | affinity, recombinant protein L (source: E. coli, sequence: amino acid 98-462 of the Peptostreptococcus Magnus) |
| Support matrix: | poly (glycidyl methacrylate-co-ethylene dimethacrylate) |
| Disk dimensions: | diameter: 12 mm; thickness: 3 mm; bed volume: 0.34 mL |
| Fitting ring: |
high density polyethylene (PE-HD); inner diameter: 12.0 mm, outer diameter: 16.0 mm |
| Dynamic binding capacity: | ≥ 12 mg IgG/mL Conditions: human IgG 1.0 mg/mL, 20 mM Tris-HCl buffer + 0.1 M NaCl, pH 7.4, flow rate 2 mL/min |
| Working flow rates: | up to 6 mL/min (320 cm/h) |
| Working system pressure: | up to 50 bar (5 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 2–10 cleaning-in-place 2–13 |
Caring for the CIM® r-Protein L Disk Monolithic Column
Regeneration
To regenerate (i.e. to wash out any strongly or unspecifically bound substances from the monolithic column), wash the column with at least 10 column volumes (3.4 mL) of a 0.1 M 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 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 following procedures can be used:
1. Removal of precipitated protein
Reverse the flow direction and:
- Wash with 5-10 column volumes of 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 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.
2. Removal of strongly bound hydrophobic proteins or lipids
Reverse the flow direction and:
- 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 the CIM® r-Protein L Disk Monolithic Column
1. Short term storage
The CIM® r-Protein L Disk Monolithic Column should be stored at +4 °C (39 °F) to +8 °C (46 °F) in buffer, e.g. Tris or PBS, in the presence of suitable bacteriostatic agent, e.g. 20 % ethanol.
2. Long term storage
The CIM® r-Protein L Disk should be thoroughly cleaned (CIP) at the end of the working day and removed from the housing. The Disk should be stored in buffer, e.g. Tris or PBS, in the presence of 20 % ethanol at +4 °C (39 °F) to +8 °C (46 °F).
