Plasmid Process Pack 1-1 mL
CIM® DEAE-1 Tube Monolithic Column Specific Information
| Color code: | green |
| Catalog number: | 100.0001-5114 |
| Tube chemistry: | weak anion exchanger; diethylamino |
| Ligand density: | 2.3 ± 0.2 mmol/g dry support |
| Support matrix: | poly (glycidyl methacrylate-co-ethylene dimethacrylate) |
| Tube dimensions: | outer diameter: 18.6 mm; inner diameter: 6.7 mm; lenght: 4.2 mm; bed volume: 1 mL |
| Connectors/ fittings: | 1/16" OD UNF 10/32 |
| Dynamic binding capacity: | ≥ 7.2 mg pDNA/mL wet support (Conditions: pDNA (4.7 kb) 0.2 mg/mL, 50 mM Tris-HCl buffer 10 mM EDTA pH 7.2, flow rate16 mL/min) |
| Working flow rates: | up to 16 mL/min (380 cm/h) |
| Working system pressure: | up to 18 bar (1.8 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 3–9 WARNING: Avoid prolonged use or use at elevated temperatures with concentrated acids (more than 0.5 M) like hydrochloric, sulphuric or acetic acid! |
Regeneration
To regenerate (to wash out from the monolithic column any ionically bound substances and to reintroduce the correct counter-ion) wash the column with at least 10 column volumes (CV) of a buffer containing 2 M NaCl at one-half of 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 completely 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 proteins
- Wash with 5–10 CV (5-10 mL) of a solution containing 1.0 M NaOH/1.0 M NaCl.
Note: Use low enough flow rates to expose the column to the cleaning reagent for several minutes. - Wash with 10 CV (10 mL) of deionized water at the working flow rate.
- Wash with 10 CV (10 mL) 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 CV (10 mL) of the working mobile phase (buffer) at the working flow rate.
2. Removal of strongly bound hydrophobic proteins or lipids
- Wash with 5–10 CV (5-10 mL) of deionized water at one-half of the working flow rate.
- Wash with 5–10 CV (5-10 mL) of a 2-propanol (30 % v/v) at one-half of the working flow rate.
- Wash with 5 CV (5 mL) of deionized water at the working flow rate.
- Re-equilibrate with 10 CV (10 mL) of the working mobile phase (buffer) at the working flow rate.
Sanitization and Storage
Sanitization of the monolithic column is achieved by thoroughly washing it with 1.0 M NaOH for at least 2 hours at room temperature.
CIM® DEAE-1 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.
CIM® C4 HLD-1 Tube Monolithic Column Specific Information
| Color code: | yellow |
| Catalog number: | 100.0001-8136 |
| Tube chemistry: | butyl |
| Ligand density: | 3.2 mmol/g dry support |
| Support matrix: | poly (butyl methacrylate-co-ethylene dimethacrylate) |
| Tube dimensions: | outer diameter: 18.6 mm; inner diameter: 6.7 mm; lenght: 4.2 mm; bed volume: 1 mL |
| Connectors/ fittings: | 1/16" OD UNF 10/32 |
| Dynamic binding capacity: | ≥ 2.5 mg pDNA/mL wet support (Conditions: pDNA (4.7 kb) 0.2 mg/mL, 50 mM Tris-HCl buffer 10 mM EDTA buffer with 3 M ammonium sulphate pH 7.2, flow rate16mL/min) |
| Working flow rates: | up to 16 mL/min (380 cm/h) |
| Working system pressure: | up to 18 bar (1.8 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 1–12 WARNING: Avoid prolonged use or use at elevated temperatures with concentrated acids (more than 0.5 M) like hydrochloric, sulphuric or acetic acid! |
Cleaning in Place (CIP)
In some cases, the simple regeneration of the monolithic column is not enough. Sample molecules may not completely 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
- Wash with 10 column volumes (10 mL) of 1.0 M NaOH.
Note: Use low enough flow rates to expose the column to the cleaning reagent for several minutes. - Wash with 10 column volumes (10 mL) of deionized water at the working flow rate.
- Wash with 10 column volumes (10 mL) of 20 % ethanol at the working flow rate.
- Re-equilibrate with 10 column volumes (10 mL) of the working mobile phase (buffer) at the working flow rate.
2. Removal of strongly bound hydrophobic proteins or lipids
- Wash with 10 column volumes (10 mL) of deionized water at one-half of the working flow rate.
- Wash with 10 column volumes (10 mL) of 2-propanol (30 % v/v) or ethanol (70 % v/v) at one-half of the working flow rate.
- Wash with 5 column volumes (5 mL) of 20 % ethanol at the working flow rate.
- Re-equilibrate with 10 column volumes (10 mL) of the working mobile phase (buffer) at the working flow rate.
Sanitization and Storage
Sanitization of the monolithic column is achieved by thoroughly washing it with 1.0 M NaOH for at least 1 hour at room temperature. CIM® C4 HLD-1 Tube Monolithic Columns 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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