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CIM® EDA-8f mL Ion Exchange Chromatography Column

 662KB

About CIM® IMAC Tube Monolithic Columns

The CIM® EDA-8f Tube Monolithic Column consists of a unique polymeric monolith encased in a specially designed housing. These columns are amine activated monolithic supports obtained by reacting the native epoxy groups with Ethylene Diamine. These columns are used for fast, highly efficient separations of large molecules such as proteins and DNA, as well as smaller molecules such as peptides. 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: 430.5116
Disk chemistry: ethylene diamino
Ligand chemistry: 3.2 ± 0.2 mmol/g dry support
Support matrix: poly (glycidyl methacrylate-co-ethylene dimethacrylate)
Tube dimensions: outer diameter: 15 mm; inner diameter: 6.5 mm; length: 56 mm; bed volume: 8.0 mL
Dynamic binding capacity: ≥ 15 mg BSA/mL wet support
(Conditions: Bovine Serum Albumin 3.0 mg/mL, 20 mM Tris-HCl buffer, pH 7.4, flow rate 16 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 50 °C (122 °F)
WARNING: Avoid prolonged use at elevated temperatures!
Recommended pH: working range 2–13
cleaning-in-place 1–14* (e. g. 1 M NaOH)
* Valid for the CIM® EDA Disk.

Caring for the CIM® IMAC-8f Tube 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 column 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 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 column volumes of a solution containing 1 M NaOH/1 M NaCl. 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.

2. Removal of strongly bound hydrophobic proteins or lipids

  • 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.

Sanitization

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

Storage of CIM® IMAC Tube Monolithic Columns

CIM® EDA-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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