CIM® CDI Disk Monolithic Column
About the CIM® CDI Disks
The CIM® CDI disks consist 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 direct coupling of proteins, peptides and other ligands containing amino or thiol groups. 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: | black with one line |
| Catalog number: | 214.8000 |
| Disk chemistry: | carbonyldiimidazole |
| Ligand density: | ∼0.6 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. WARNING: Polyoxymethylene is not stable in concentrated organic solvents, strong acids, or oxidizing agents, especially at elevated temperatures! |
| 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 1–14 cleaning-in-place 1–14 (e. g. 1 M NaOH) WARNING: The working range is only valid for the matrix itself. After coupling, it depends on the stability of the coupled ligand! |
Coupling on the CIM® CDI Disk Monolithic Columns
Coupling the ligand
Coupling is performed at a higher pH where the CDI groups are more reactive. The upper pH value is limited by the ligand stability.Since this is specific for any particular ligand, it should be thoroughly checked and optimized from case to case. In our experience, coupling at pH 8–11 is a good compromise.
Temperature dependence
The coupling of the ligand to the CIM® CDI Disk is obtained by a covalent reaction between the CDI groups on the support and the amino or thiol groups of the ligand. Therefore, the coupling rate increases exponentially with temperature according to the Arrhenius equation. From this point of view, an elevated temperature would be preferred for shortening the coupling time. On the other hand, many ligands may become unstable at higher temperatures and tend to agglomerate or even to lose their biological activity. The coupling temperature depends on the stability of your ligand and should be optimized from case to case. For most ligands (especialy those with high molecular masses) the coupling is best performed for several days at 4°C. If, however, the ligand is stable, room or even higher temperature is recommended.Buffer composition
According to our experience, the buffer type does not significantly influence the coupling efficiency. Of course, in some cases, the selection of the buffer is of utmost importance, especially when the ligand stability depends on the environment. Because of this, physiologic buffers are preferred. For coupling, the ligand should be dissolved in a coupling buffer. In some cases, it is necessary to add organic solvents to dissolve the ligand. The polymeric backbone of the CIM® Disk is hydrophobic. Because of this, coupling can be enhanced by using buffers with higher ionic strengths (e. g. 0.5 M). In this way, the ligand comes closer to the surface facilitating coupling. Higher ionic strengths may not be appropriate as some ligands can start to agglomerate and hinder coupling.Blocking the excess reactive groups
Typically after immobilization the remaining reactive groups must be blocked in order to avoid unspecific interactions during use. However, with the CDI chemistry blocking excessive reactive groups is not necessary since the groups left are hydrolyzed into OH groups which are not problematic.Storage of CIM® CDI Disk Monolithic Columns
Unused CIM® CDI Disk Monolithic Columns should be stored in 96 % ethanol solution at a temperature of +4 °C to +8 °C. Used CIM® CDI Disk Monolithic Columns (immobilized with a ligand) should be stored in conditions that suite the ligand in question.LOOK-UP CHEMISTRY
Select chemistry from the list:
On-line CATALOG
BROCHURES
 |
 |