PRODUCTS > CIM® Laboratory Columns

CIM® DEAE Disk Monolithic Column

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About the CIM® DEAE Disks

The CIM® DEAE Disk consists of a unique monolithic stationary phase that is easily placed in a dedicated plastic housing forming a CIM® Disk Monolithic Column. These columns are used for fast, highly efficient separations of large molecules such as proteins, DNA or viruses, as well as smaller molecules such as peptides. 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: green
Catalog number: 210.5114
Disk chemistry: weak anion exchanger; diethylamino
Ligand density: 2.3 ± 0.2 mmol/g dry support
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 mm, outer diameter: 16 mm
Dynamic binding capacity: ≥ 21 mg BSA/mL wet support
(Conditions: Bovine Serum Albumin 1.0 mg/mL, 20 mM Tris-HCl buffer, pH 7.4, flow rate 3 mL/min)
Working flow rates: up to 10 mL/min (530 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 50 °C (122 °F)
WARNING: Avoid prolonged use at elevated temperatures!
Recommended pH: working range      3–9
cleaning-in-place 2–14 (e. g. 1 M NaOH)
WARNING: Avoid prolonged use or use at elevated temperatures of CIM® DEAE disk with concentrated acids (more than 0.5 M) like hydrochloric, sulphuric or acetic acid!

Caring for the CIM® DEAE Disk Monolithic Columns

Regeneration

To regenerate (to wash out from the monolithic column any ionically bound substances and to reintroduce the correct counter-ion) wash the disk with at least 10 column volumes 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 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 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 5–10 column volumes of a solution containing 1.0 M NaOH/1.0 M NaCl.
    Note: Reverse the flow direction and use low enough flow rates to expose the column to the cleaning reagent for several minutes.
  • Wash with 10 column volumes of distilled 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.
2. Removal of strongly bound hydrophobic proteins or lipids
Reverse the flow direction and:
  • Wash with 5–10 column volumes of distilled 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 distilled water at the working flow rate.
  • Re-equilibrate with 10 column volumes of the working mobile phase (buffer) at the working flow rate.

Sanitization

Sanitization of the monolithic column is obtained by thoroughly washing it with 1.0 M NaOH for at least 1 hour at room temperature.

Sterilization

Sterilization of the CIM® DEAE disk can be obtained by autoclaving the monolith at 121 °C for 15 min.
WARNING: You can autoclave the disk itself, or the combination of a disk and the PEEK (grey) housing. Never autoclave the combination of a disk and the polyacetal (white and blue) housing.
CAUTION: With the anion exchangers, it is posible that a progressive degradation of the tertiary and quaternary amine groups can occur.

Storage of the CIM® DEAE Disk Monolithic Columns

1. Short term storage
The CIM® DEAE Disk Monolithic Columns should be stored at a temperature of +4 °C to +8 °C in the presence of a suitable bacteriostatic agent, e. g. 20 % ethanol.

2. Long term storage
CIM® DEAE Disk should be thoroughly cleaned (CIP) at the end of the working day and removed from the housing. The disk should be stored in a jar with 20 % ethanol.

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