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PRODUCTS > CIM^® Laboratory Columns

CIM^® IDA Disk Monolithic Column (PEEK Housing)

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About the CIM^® IDA Disks

The CIM^® IDA Disk consists of a unique monolithic stationary phase that is easily placed in a dedicated plastic housing forming a CIM^® Disk Monolithic Column. These affinity columns are used for the fast, highly efficient purification of histidine containing or (His)-tagged proteins, which rely on histidines' affinity for immobilized transition metals. The short monolithic length and the specially engineered highly porous structure allow operations at elevated flow rates with low-pressure drops (only a few bar) while preserving the binding capacity and resolution.
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:	orange
Catalogue number:	217.3010
Disk chemistry:	affinity, iminodiacetic acid (IDA) as chelating ligand (uncharged)
Support matrix:	poly (glycidyl methacrylate-co-ethylene dimethacrylate-co-ethyl methacrylate)
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
Metal ion capacity:	23 ± 10 μmol M2+/mL of support (metal ion dependent)
Housing:	PEEK (Catalog #: 222.0850)
Working flow rates:	up to 8 mL/min (420 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:	pH>3

Caring for the CIM^® IDA Disk Monolithic Columns

Charging

When the CIM^® IDA disk monolithic column is to be charged for the first time, the following procedure should be performed:

• Wash with 20 column volumes of distilled water
• Charge with 10 column volumes of 30 mM metal salt solution (e. g. CuSO4, NiSO4)
• Wash with 20 column volumes of distilled water
• Re-equilibrate with 20 column volumes of the working mobile phase (buffer) at the working flow rate.

Regeneration

When the CIM^® IDA disk monolithic column is to be recharged with metal ions, the following procedure should be performed:

• Wash with 20 column volumes of distilled water
• Wash with 10 column volumes of 1 M HCl to strip the metal ions
• Wash with 20 column volumes of 0.5 M phosphate buffer/ 1 M NaCl, pH 7.4
• Wash with 20 column volumes of distilled water
• Recharge with 10 column volumes of 30 mM metal salt solution (e. g. CuSO4, NiSO4)
• Wash with 20 column volumes of distilled water
• Re-equilibrate with 20 column volumes of the working mobile phase (buffer) at the working flow rate.

Cleaning In Place (CIP)

The disk monolithic column does not have to be cleaned between purifications if the same protein is to be purified. It is sufficient to clean the disk monolithic column after 3–5 purifications, depending on the cell lysate, target proteins, conditions used, etc. However, if an increase in back pressure is seen then the CIM^® disk monolithic column must be cleaned. The following procedures can be applied:
WARNING: The columns must be regenerated after these CIP procedures are performed.

1. Removal of precipitated protein:
   • Wash with 20 column volumes of distilled water
   • Wash with 20 column volumes of 1 M NaOH
   • Incubate with 1 column volume of NaOH for 1 hour
   • Wash with 20 column volumes of 0.5 M phosphate buffer/ 1 M NaCl, pH 7.4
2. Removal of strongly bound hydrophobic proteins or lipids
   • Wash with 20 column volumes of distilled water at one-half of the working flow rate.
   • Wash with 20 column volumes of a 30 % 2-propanol at one-half of the working flow rate.
   • Wash with 20 column volumes of distilled water at the working flow rate.
   • Re-equilibrate with 20 column volumes of the working mobile phase (buffer) at the working flow rate.

Reducing nonspecific binding

• High concentrations of NaCl (1–2 M) are recommended in the binding, washing and elution buffer.
• Low concentrations of imidazole (1–20 mM) in the lysis or binding and washing buffer are recommended.
• Detergent (Tween 20 or Triton X) up to 2 % can be used.
• Use of deoxyribonuclease (DNase) and ribonuclease (RNase) in the cell lysis procedure is recommended.

Storage of the CIM^® IDA Disk Monolithic Columns

1. Short term storage
The CIM^® IDA 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
The CIM^® IDA 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.