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PRODUCTS > CIM^® Laboratory Columns

CIM^® r-Protein G Disk Monolithic Column

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About the CIM^® r-Protein G Disks

The CIM^® r-Protein G 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 fast, highly efficient purification of immunoglobulin G.
r-Protein G binds the Fc portion of antibodies (IgG class) without affecting antigen binding. In comparison to protein A, the range of polyclonal IgGs that bind strongly to r-Protein G is much wider and adsorption takes place over a wide pH range. 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:	salmon
Catalogue number:	217.1011
Disk chemistry:	affinity, r-Protein G from Streptococcus aureus
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 (HD-PE); inner diameter: 12 mm, outer diameter: 16 mm
Dynamic binding capacity:	≥ 10 mg/mL (Conditions: human IgG 1.0 mg/mL, 20 mM Tris-HCl buffer, pH 7.4, flow rate 3 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        2–11 cleaning-in-place  2–13

Caring for the CIM^® r-Protein G Disk Monolithic Columns

Regeneration

To regenerate (to wash out from the monolithic column any strong or unspecifically bound substances), wash the disk with at least 10 column volumes of a 0.1 M buffer containing 1.0 M NaCl, pH 7.0–8.0 at one-half of the working flow rate. After that, wash the disk 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 disk 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 proteins

• Wash with 5–10 column volumes of a 0.1 M NaOH.
  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.

Storage of the CIM^® r-Protein G Disk Monolithic columns

1. Short term storage
The CIM^® r-Protein G 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^® r-Protein G 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.