Aether SP-650

Aether SP-650 IEX resin is based on polyacrylate; the surface is hydrophilically modified then bonded with strong acidic ion-exchange groups. It has high loading capacity, good chemical stability, high mechanical strength and small non-specific adsorption. It features excellent biocompatibility and column bed stability, and can provide faster flow rate. It is especially suitable for large-scale preparation applications, significantly improving the production efficiency of downstream purification process, reducing costs and creating better economic benefits. It is widely used in capture, intermediate purification and final polishing of antibodies, proteins, peptides, nucleic acids (oligonucleotides), viruses, insulins and other biomolecules.

Method of use

Chromatography steps

1. Equilibration. Equilibrate the column with 5–10 CV of a suitable buffer until the conductivity and pH of the eluent are stable and consistent with the equilibration buffer. For example, the equilibration buffer could be 20 mM PBS, pH 7.0. Screen and optimise according to the stability and isoelectric point of the target protein and the type of ion-exchange resin.
2. Sample loading. Prepare solid samples by dissolving them in equilibration buffer. Dialyse low-concentration sample solutions against equilibration buffer; dilute high-concentration sample solutions with equilibration buffer. To avoid column clogging, centrifuge or membrane filter the sample (preferably 0.45 or 0.22 µm). Calculate the amount of feed from the loading capacity of the resin and the content of the target protein in the feed solution. Before loading, make the sample buffer as consistent as possible with the equilibration buffer.
3. Elution. After loading, continue to rinse with equilibration buffer until the baseline is stable. Depending on the separation, elute the adsorbed samples in sequence by increasing the salt concentration or changing the pH of the mobile phase.
4. Regeneration. After each chromatography run, wash the column with 0.5–2M NaCl to remove proteins strongly bound to the resin.

Cleaning and regeneration

Contaminants such as lipids, endotoxins and proteins accumulate on the column as the number of uses increases. Determine the frequency of cleaning-in-place (CIP) according to the degree of contamination. Where contamination is considerable, CIP after each use is recommended to ensure repeatability of results and to prolong the working life of the resin.

Recommended cleaning conditions by contaminant type:

• Strongly binding proteins: wash with 5 CV of 2M NaCl solution, or use a high-salt buffer not lower than pH 2, such as 1M NaAc solution.
• Strongly hydrophobic and precipitated proteins: wash with 5 CV of 0.2–0.5M NaOH solution (contact time 1–2 hours), then wash out the alkali with 5–10 CV of ultrapure or pure water.
• Lipoproteins and lipids: wash with 5 CV of 50% ethanol or 30% isopropanol (contact time 0.5–1 hour), then rinse with 5–10 CV of ultrapure or pure water. Alternatively, clean with an alkaline or acidic solution containing a non-ionic surfactant, such as 0.1–0.5% Triton X-100 + 0.1M acetic acid for 12 hours, then rinse with more than 5 CV of 50% ethanol to remove the detergent, followed by 5 CV of ultrapure or pure water. When using high-concentration organic solvents, increase the solvent concentration gradually to avoid air bubbles.

Degas 50% ethanol or 30% isopropanol before use. Use a flow rate of 30–60 cm/h during CIP. Use reverse cleaning where clogging is severe.

To reduce microbial load, treat the resin with 0.5–1M NaOH solution.

Storage

Keep unopened resin in the original container at 4–30°C in a well-ventilated, dry and clean place. Do not freeze. Wash the used column with 2–3 CV of 20% ethanol solution and store at 2–8°C.

Destruction and recycling

Chromatography resin is difficult to degrade in nature, so incineration of waste resin is recommended. For resin that has been in contact with biologically active samples such as viruses and blood, follow local biosafety requirements before destroying or disposing of it.

Packing method

Detailed information on resin packaging is available on request. Please contact your local distributor.

Ordering information

• 228-00025 — 25 ml
• 228-00100 — 100 ml
• 228-00500 — 500 ml
• 228-01000 — 1 L
• 228-05000 — 5 L
• 228-10000 — 10 L