Persecryl S-100

Persecryl S-100 is a hydrophilic gel filtration SEC resin with a particle size of 25–75 µm and a separation range of 1–100 kDa. It gives a high flow rate in column operation, with high resolution and recovery. The resin is used in the separation and purification of biological macromolecules such as enzymes, polysaccharides, nucleic acids and proteins, and in the production of biological products including interferon-γ, interleukin 2, protein A and hepatitis B vaccine.

Method of use

Chromatographic conditions

Buffer selection. Consider the stability of the sample in the buffer. Use a salt-containing buffer rather than ultrapure or pure water, to avoid non-specific adsorption.

Flow rate. Select a linear flow rate no higher than 125 cm/h according to the column bed height.

Sample pretreatment. Filter the sample through a 0.45 µm microporous membrane before loading to prevent the column from clogging.

Chromatography steps

1. Equilibration. Use the buffer to fully equilibrate the column until the pH and conductivity are stable and consistent with the equilibration buffer. This step usually requires 2–5 column bed volumes (CV).
2. Sample loading. The usual loading volume is 1–2% of the column volume. Do not use too high a sample concentration, to avoid overpressure and loss of resolution.
3. Elution. Elute with buffer and collect the peaks at different positions, usually over 1–1.5 CV.
4. Regeneration. Rinse the column with a high-salt buffer, such as 1M NaCl.
5. Re-equilibration. Re-equilibrate the column with buffer.

Cleaning and regeneration

Contaminants such as lipids, endotoxins and proteins accumulate on the column as the number of uses increases. Regular cleaning-in-place (CIP) keeps the column in a stable working condition. Determine the frequency of 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 1–2 CV of 2M NaCl solution.
• Strongly hydrophobic and precipitated proteins: wash with 1 CV of 0.1M NaOH solution, then wash out the alkali with 5–10 CV of ultrapure or pure water.
• Lipoproteins and lipids: wash with 5 CV of 70% ethanol or 30% isopropanol, then rinse with 5–10 CV of ultrapure or pure water.

Degas 70% ethanol or 30% isopropanol before use. Keep the flow rate at no more than 60 cm/h during CIP. Reverse cleaning can be used where clogging is severe.

After cleaning, equilibrate the column with at least 3 CV of equilibration buffer, until the pH and conductivity remain unchanged.

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

• 201-00025 — 25 ml
• 201-00100 — 100 ml
• 201-00500 — 500 ml
• 201-01000 — 1 L
• 201-05000 — 5 L
• 201-10000 — 10 L

‍