Oligonucleotide Purification
Hamilton’s PRP-C18 is the First Choice for Oligonucleotide Purifications
Synthetic oligonucleotide (oligo) purifications are demanding applications that are typically performed at elevated (> 60°C) temperatures in order to facilitate resolution between the target oligo and closely-related failure sequences. Traditional silica-based C18 (ODS) columns have diminished lifetime at elevated temperatures, however, polymeric polystyrene-divinylbenzene (PS-DVB) columns are routinely used to purify synthetic oligonucleotides due to their superior temperature (> 100°C) and pH (1–14) stability.
Why PRP-C18?
The Hamilton PRP-C18 is a next generation polymeric column based on porous C18-functionalized PS-DVB that exhibits excellent chemical and thermal stability. Unlike other polymer columns, the octadecylated surface modification imparts superior mass transfer kinetics for high efficiency separations and excellent mechanical stability.
PRP-C18 Benefits:
- Superior resolution for purer oligos
- Extended column lifetime for increased productivity
- High loading capacity for time and cost savings
Resilient Design Ensures More Oligo Manufacturing
The flexibility to employ elevated temperatures (up to 100°C) and alkaline pH are important tools in oligonucleotide purifications. Traditional silica-based supports break down at elevated temperatures, leading to stationary phase bleeding and diminished column lifetimes. The PRP-C18 does not dissolve, phase-strip, or bleed even under the most extreme operating conditions, unlocking the power of superior resolution between long oligos and their failure sequences. Because the PRP-C18 stationary phase is devoid of silanols found in silica-C18 columns, peak tailing, poor recovery, and carry-over problems are completely eliminated, offering significant value.
Full pH Range Compatibility
Unlike traditional silica-based C18 columns, the PRP-C18 is stable over a pH range of 1–14. The flexibility to employ alkaline mobile phase (pH 12) is useful for separating problematic oligos that form secondary structures or aggregates.
High pH Column Lifetime Comparison
Forced degradation study comparing longevity of traditional ODS and Hamilton PRP-C18 columns under elevated temperature (60°C) and alkaline pH (50 mM phosphate, pH 12) conditions.
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Higher Oligo Yield with Extended Loading Capacity
The high loading capacity and greater resolving power of the PRP-C18 makes it possible to achieve baseline resolution between failure sequences and the target 21mer, which is collected in a single fraction. Subsequent analytical chromatography of the purified fraction indicate oligo purification to > 99% (UV).
2 mg prep load
10 x 100 mm, 5 μm PRP-C18
Confirmed > 99% purity
4.1 x 50 mm, PRP-C18
High Purity Oligos with High-Resolution Separations
The performance of the PRP-C18 and two leading oligo columns at increasing temperatures is shown in the graphs below. The PRP-C18 shows the best resolution of poly(dC) 12–18 ladder at all temperatures.
Waters XTerra™
2.1 x 100 mm, 5 μm
Hamilton PRP-C18
2.1 x 150 mm, 5 μm
Ordering Information
* Not Seeing the Hardware Dimensions you need? We offer many custom sizes to suit your needs.
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PRP-C18 Guard Columns Ordering Information
| Part Number | Description |
|---|---|
| 79685 | Analytical Guard Column Starter Kit (1 holder, 2 cartridges), Stainless Steel |
| 79686 | Analytical Guard Column Replacement Cartridges (5/pk), Stainless Steel |
| 79687 | Analytical Guard Column Starter Kit (1 holder, 2 cartridges), PEEK |
| 79688 | Analytical Guard Column Replacement Cartridges (5/pk), PEEK |
| 79689 | Semi-prep/Preparative Guard Column Starter Kit (1 holder, 1 cartridge), Stainless Steel |
| 79690 | Semi-prep/Preparative Guard Column Replacement Kit (2/pk), Stainless Steel |
Other Packing Materials for Oligo Purification
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