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CIMmultus® DEAE columns are weak anion exchange monoliths designed for fast, gentle, and scalable purification of negatively charged biomolecules – especially plasmid DNA (pDNA). DEAE provides a milder, pH tunable interaction than strong AEX, helping you bind at near neutral pH and elute under controlled, product friendly conditions. Combined with the convective flow of CIMmultus monoliths, DEAE enables short cycle times, low backpressure, and straightforward transfer from development to manufacturing.
CIMmultus® DEAE: Weak Anion Exchanger for pDNA Capture
CIMmultus® DEAE columns employ diethylaminoethyl (DEAE) as a ligand, a weak-base tertiary amine that is positively charged when protonated. This ligand binds molecules with predominantly negative charge and repels molecules with a predominantly positive charge. DEAE column is typically used for first capture step of pDNA purification, where contaminant RNA and proteins are separated from plasmid DNA. The purification process involves binding of negatively charged solutes (e.g., plasmid DNA) under suitable pH and conductivity, followed by elution by increasing ionic strength, such as NaCl gradients.
Chromatography Mode: Weak anion exchange (AEX) with DEAE ligand
Ligand Density: 0.45 – 0.65 mmol/mL wet support
Dynamic Binding Capacity:
2 µm channel size: 4 – 6 mg of pDNA/mL of monolith
6 µm channel size: up to 2.5 of pDNA/mL of monolith
pH ranges: Operation: pH 2–10 | Cleaning in place (CIP): pH 1–14
Operating Flow Rate and Maximum Pressure: Refer to Instructions for Use (IFU) for your column volume
Chemical Compatibility: Compatible with commonly used aqueous buffers, 1 M NaOH, 0.1 M HCl, 8 M urea, 6 M guanidine hydrochloride, and 20% ethanol
Cleaning and Sanitization
CIP: 0.5 M NaOH + 2 M NaCl
Neutralization | Equilibration: Concentrated buffer (e.g., >100 mM Tris, pH ~7) with high salt (e.g., >1 M NaCl) or 1 M ammonium acetate
Storage: 20% ethanol
Shelf Life: 7 years
Choosing the right channel size depends on the size of your pDNA. We suggest using the CIMmultus DEAE column with channels having 2 μm of diameter if the plasmid has less than 8 kbp, whereas if the plasmid is larger and has more than 8 kbp then choosing 6 μm channels is a better option. Larger plasmids generate more viscous solutions, sometimes limiting the operating flow rates. In addition, open-circular isoforms of large plasmid DNA can become reversibly entrapped in the chromatographic column. Large channel (6 μm) monoliths successfully prevent pressure increase and entrapment of oc isoform. The 1.3 μm channel diameter is not recommended for plasmids, it is usually used for anion exchange chromatography of proteins which are generally smaller than pDNA.
Yes, monolithic columns are designed for multiple uses, their lifetime depends on the sample and column maintenance. It is recommended to clean the column (at least 2h in cleaning solution) after every purification run. We recommend following the instructions written in Instructions for Use and optimize the cleaning for your sample.
When equilibrating the DEAE column it is important to pre-expose the column to both eluting and binding environment. Equilibration is performed by washing the column with at least 10 CVs of elution buffer (which contains a higher concentration of salt), followed by washing with a buffer that matches the binding conditions to prepare the column (50 mM TRIS, 10 mM EDTA, pH 7.2).
We do not recommend loading crude lysate on the column. Unfiltered samples can foul the column and increase backpressure on the column. Impurities will also compete with binding sites on the column and lower its capacity, and product yields.
