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 anion exchange resins use polymethacrylate as the matrix. The hydrophilicity of the matrix is improved through surface modification. Meanwhile, through proprietary nano-micro structure regulation technology, products with different pore sizes ranging from 30-1000nm can be achieved. It fully meets the separation and purification needs of biological macromolecules including peptides, recombinant proteins, nucleic acids, vaccines, antibodies, viruses, and virus-like particles. High-throughput capacity is also suitable for viscous loading samples. Products with different sizes and pore sizes are available according to application needs, meeting the requirements of capture, intermediate purification, polishing, and analytical purification processes. Duoning provides comprehensive solutions for biological sample purification from laboratory to industrial production.

2.Features

Table 1: Duoning Small-pore Anion Exchange Resins HPQ series

Table 2: Duoning Large-pore Anion Exchange Resins CPQ series

Table 3: Duoning Macroporous Anion Exchange Resins GPQ series

Note:

1. The mobile phase pH must be between the PI and PKa values; for pH > 10.0, strong anion series are recommended.

2. For special particle size requirements, professional customized services is available.

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.