1.Details

Ion Exchange Chromatography (IEC) is the most commonly used method for the separation and purification of biomacromolecules based on the type, number, and distribution of its specific surface charges. Q HL FF anion exchange resins uses agarose as the matrix，high loading quaternary ammonium group as ligand. Duoning Q HL FF has the characteristics of high binding capacity and easy scalability and can meet the application requirements of capture, medium purification, polishing, and analytical purification in various stages. Duoning provides comprehensive solutions for biological sample purification from laboratory to industrial production.

2.Features

Table 1: Duoning high-load Anion Exchange Resins QHL FF series

Protocol

Ion exchange resin is widely used for the separation and purification of proteins, plasma products, enzymes, polysaccharides, nucleic acids, etc. Chromatographic operation typically includes steps such as equilibration, sample loading, washing, elution, and regeneration. The specific operating methods are as follows:

Alkaline wash: Rinse the chromatographic column with 0.5 M NaOH at a low flow rate for 3 column volumes (CV).

Equilibration: Equilibrate the column with 5-10 CV of equilibration buffer (e.g., 20 mM PB, pH 7.0; the specific buffer system should be selected and optimized based on the stability and isoelectric point of the target protein) at a flow rate not exceeding the maximum pressure limit, until the conductivity and pH of the effluent remain constant (consistent with the equilibration buffer).

Sample Loading: The sample loading buffer should be as consistent as possible with the equilibration buffer; solid samples can be dissolved and prepared in equilibration buffer; dilute samples can be dialyzed with equilibration buffer or supplemented with the corresponding amount of salt; concentrated sample solutions can be diluted with equilibration buffer. To avoid clogging the chromatography column, sample solutions should be centrifuged or microfiltered (0.45 μm). The loading volume should be calculated based on the binding capacity and the target protein content in the sample solution.

Washing: After loading, continue washing with equilibration buffer until the UV absorbance drops to baseline.

Elution: Elute with elution buffer (e.g., 20 mM PB + 1M NaCl, pH 7.0). Elution methods include pH gradient elution, linear gradient elution, or step gradient elution.

Regeneration: After each cycle, clean the column with 1-2 M NaCl to remove strongly bound proteins.

Cleaning In Place (CIP): After 5-10 uses (the specific number depends on the type and source of raw materials and experimental requirements), the column requires Cleaning In Place, which can refer to the table below.

Storage: Store in 20% ethanol at 2-30°C; The unpacked resins can be flushed with 20% ethanol and then stored at 2-30°C.

Other Considerations: When using and storing the column, avoid letting it run dry or become poorly sealed to prevent air bubbles from entering.
3.Specification

Duoning Biotech is a leading one-stop bioprocess solutions provider from China, committed to offering products and services required for the discovery, development and commercialization production of biologics, including reagents, consumables, instruments, equipment, and services. Leveraging our integrated bioprocess system, we empower our customers to achieve effective, reliable, lower-cost and higher-quality biologics manufacturing processes.