Universal Linker "Maleimide-Type Linker Supports"
In solid-phase synthesis using the phosphoramidite method, nucleoside linkers--nucleosides fixed to a solid support via a linker--are used to attach the 3' terminal nucleoside of the oligonucleotide being synthesized. Since the 3' terminal nucleoside varies depending on the oligonucleotide sequence, this method requires preparing eight types of nucleoside linkers for DNA (dA, dG, dC, dT) and RNA (rA, rG, rC, rU), as well as additional modified nucleoside linkers when synthesizing modified oligonucleotides, which can be cumbersome. To solve this problem, solid supports modified with a universal linker have been developed as an alternative to conventional nucleoside linkers.
Currently, the most widely used universal linker is UnyLinker™*, developed by Ionis Pharmaceuticals, which is a linker bearing a maleimide ring. In recent years, universal linkers with new backbones, as well as conditions for cleaving the universal linker from the solid support, have also been developed.
This product is a solid support modified with a universal linker developed by Ionis Pharmaceuticals. It is offered under the product name "Maleimide-type Linker." A lineup of multiple pore sizes is available, so you can choose the optimal pore size according to the type of oligonucleotide to be synthesized.
*UnyLinker™ is a trademark of Ionis Pharmaceuticals, Inc.
Applications
Synthesis of oligonucleotides (T10 and T20) using Maleimide-Type Linker PS in DMTr-Off mode
1) Synthesis of T10mer
| Solid Supports | UnyLinker™ | Crude Yield (%) |
Crude Purity (HPLC, area%) |
|||||
|---|---|---|---|---|---|---|---|---|
| Supplier | Materials | Pore Size (Å) | Substituent on N | Functional Loading (µmol/g) |
T9 <N-1> |
T10 <FLP> |
||
| 1 | FUJIFILM Wako | HCP (Low-Swelling Polymer) |
1000 | Ph | 49 | >90 | 5 | 92 |
| 2 | 2000 | 27 | >90 | 3 | 92 | |||
| 3 | 3000 | 29 | >90 | 4 | 90 | |||
| 4 | 4000 | 8 | >90 | 4 | 90 | |||
| C1 | Competitor A | CPG | 500 | Ph | 90 | 69 | 10 | 83 |
| C2 | 1000 | 48 | 71 | 7 | 87 | |||
| C3 | Competitor B | CPG | 1000 | Me | 45 | 67 | 2 | 93 |
| C4 | Competitor C | Swelling Polymer | Swelling Pore | Ph | 407 | 73 | 1 | 95 |
| C5 | Competitor D | 347 | 79 | 2 | 94 | |||
Note: These results were obtained from limited sample sizes and specific conditions
a) FUJIFILM Wako PS 1000Å
b) Competitor CPG 1000Å
Figure HPLC analysis of ONs released from T10-loaded (a) UnyLinker™ PS 1000Å (R=Ph) and (b) UnyLinker™ CPG 1000Å (R=Me) under Basic conditions. Basic condition: AMA at 60℃ for 1 h.
HPLC condition of Oligonucleotide
| Column size | : | Wakopak® Ultra C18-2 φ2.1 mm×100 mm (D) |
| Mobile phase | : | A) 100 mM TEAA aq., B) Acetonitrile |
| Gradient | : | 0-15 min B=5-15%, 15-20 min B=100%, 20-25 min B=5% |
| Flow rate | : | 0.3 mL/min |
| Temperature | : | 40℃ |
| Detection | : | UV260 nm |
| Injection vol. | : | 0.5 µL |
| Sample | : | DNA dT 10 mer, All PO |
2) Synthesis of T20mer
| Solid Supports | UnyLinker™ | Crude Yield (%) |
Crude Purity (HPLC, area%) |
|||||
|---|---|---|---|---|---|---|---|---|
| Supplier | Materials | Pore Size (Å) | Substituent on N | Functional Loading (µmol/g) |
T19 <N-1> |
T20 <FLP> |
||
| 1 | FUJIFILM Wako | HCP (Low-Swelling Polymer) |
1000 | Ph | 29 | >90 | 5 | 87 |
| 2 | 2000 | 19 | >90 | 4 | 83 | |||
| 3 | 3000 | 20 | >90 | 5 | 80 | |||
| 4 | 4000 | 8 | >90 | 5 | 84 | |||
| C1 | Competitor A | CPG | 500 | Ph | 90 | 73 | 12 | 80 |
| C2 | 1000 | 48 | 69 | 7 | 83 | |||
| C3 | Competitor B | CPG | 1000 | Me | 45 | 68 | 8 | 86 |
| C4 | Competitor C | Swelling Polymer | Swelling pore | Ph | 407 | 57 | 4 | 79 |
| C5 | Competitor D | 347 | 69 | 12 | 78 | |||
Note: These results were obtained from limited sample sizes and specific conditions
HPLC condition of Oligonucleotide
| Column size | : | Wakopak® Ultra C18-2 φ2.1 mm×100 mm (D) |
| Mobile phase | : | A) 100 mM HFIP + 8.6mM TEA aq., B) Acetonitrile |
| Gradient | : | 0-15 min B=1-9%, 15-20 min B=100%, 20-25 min B=1% |
| Flow rate | : | 0.3 mL/min |
| Temperature | : | 40℃ |
| Detection | : | UV260 nm |
| Injection vol. | : | 0.25 µL |
| Sample | : | DNA dT 20 mer, All PO |
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