DEAE-60 Weak Cation Exchange Medium

Product Introduction:

Ion exchange chromatography is the most commonly used method for the separation and purification of biomacromolecules. The HPDEAE-60 weak anion exchange medium developed by Bojin Biotechnology uses polymethacrylate microspheres with rigid structure as the matrix, and uses a unique surface modification technology to greatly improve the hydrophilicity of the polymer matrix. Heterotropic adsorption and excellent biocompatibility and other advantages. The designed pore size is widely applicable to the purification of most antibodies and recombinant proteins except polypeptides and viruses.

Application

DEAE-60 Weak Cation Exchange Medium is specially designed for the separation and purification of recombinant proteins. Its chromatographic operation usually includes steps such as equilibration, sample loading, rinsing, elution, and regeneration.

The specific operation method is as follows (taking 1ml column as an example)

1. Equilibrium: Use 5~10CV of equilibrium buffer (Buffer A, such as 20 mM PB, pH7.0, the specific buffer system should be screened and optimized according to the stability and isoelectric point of the target protein) at a pressure not higher than the upper limit pressure. Balance the ion exchange column at a flow rate until the conductance and pH of the effluent remain unchanged (consistent with the equilibrium solution).
2. Sample loading: The sample buffer solution should be as consistent as possible with the balance solution. Solid samples can be prepared by dissolving the balance solution; low-concentration sample solutions can be dialyzed with the balance solution or a corresponding amount of salt can be added; high-concentration sample solutions can be diluted with the balance solution. To avoid clogging the column, samples should be processed by centrifugation or microfiltration (0.45 μm). The amount of feed is calculated according to the loading capacity of the medium and the content of the target protein in the feed liquid.
3. Rinse: Continue to rinse with equilibration buffer to baseline.
4. Elution: Elute with elution buffer (20 mM PB+1M NaCl, pH 7.0, pH gradient elution can also be used) (linear gradient elution or step gradient elution can be used), and the effluent is collected.
5. Regeneration: After each chromatography, the column can be washed with 1-2M NaCl to remove strongly bound proteins.
6. In-situ cleaning: After the medium is used 5-10 times (the specific number is related to the type and source of the raw material and the experimental requirements), the medium needs to be cleaned in situ:

               1. For proteins that are strongly bound by ionic bonds, wash with 2M NaCl for 3~4CV

               2. For precipitated proteins, hydrophobically bound proteins, and lipoproteins, wash with 1M NaOH for 3~4CV

               3. For proteins, lipoproteins and lipids with strong hydrophobic binding, wash with 70% ethanol or 30% isopropanol at  a flow rate not higher than the upper limit

                   pressure for 3~4CV