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Solid Phase Synthesis Technology for Nucleic Acid Drugs

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Solid-phase synthesis of nucleic acid drugs is a crucial method in biological sciences. CD Formulation provides advanced automated synthesis equipment and an experienced technical team to create high-purity, high-yield nucleic acid products customized to customers' needs.

Advantages of Our Solid Phase Synthesis Technology for Nucleic Acid Drugs

Solid-phase synthesis involves gradually adding nucleic acid bases to a solid carrier to extend the molecule's length through chemical reactions. This method enables the synthesis of nucleic acid molecules with specific sequences by immobilizing the 3'-end of the chain on a solid carrier and adding bases sequentially. The molecules are released from the carrier after each reaction by dissolving it, resulting in the synthesis of nucleic acid molecules with desired sequences.

Fig.1 Solid phase synthesis technology advantages for nucleic acid drugs Fig.1 Advantages of nucleic acid drugs solid phase synthesis technology. (CD Formulation)

Carriers for Solid Phase Synthesis of Nucleic Acid Drugs

CD Formulation utilizes various solid-phase carriers for nucleic acid solid-phase synthesis. These carriers are chemically stable and mechanically robust, effectively immobilizing the first base monomer and supporting subsequent synthesis steps based on specific nucleic acid requirements and customer applications.

Common solid-phase carriers for nucleic acid synthesis have specific characteristics.

Solid-phase Carriers	Features	Advantages
Controlled Pore Glass (CPG)	CPG exhibits excellent chemical resistance, high mechanical strength, and controlled pore size.	It is suitable for synthesizing long-chain nucleic acids, with high elution efficiency and uniform pore size distribution, promoting homogeneous reactions.
Polystyrene (PS)	High chemical inertness, good mechanical strength, and moderate pricing.	Widely used in various biosynthesis reactions, easy to handle, and suitable for large-scale synthesis.
Polyethylene Glycol (PEG)	The structure of the chain is flexible, with good solubility and biocompatibility.	It can enhance the yield and purity of nucleic acid chains, minimize side reactions, and is particularly suitable for nucleic acid synthesis with special modifications and intracellular applications.
Monomolecular Glass (MMG)	Monodispersed glass substrate.	Provides a high-purity synthesis environment to reduce non-specific reactions and strand breaks.

Our Workflow for Solid Phase Synthesis of Nucleic Acid Drugs

CD Formulation specializes in nucleic acid drug development and manufacturing, with expertise in solid-phase synthesis. The process includes four main steps: deprotection, coupling, oxidation, and capping.

Fig.2 Phosphoramidite chemistry for the synthesis of oligonucleotides in solid phase Fig.2 Solid-phase synthesis of oligonucleotides by phosphoramidite chemistry. (Bege M, et al., 2021)

De-protection

This step is essential in solid-phase synthesis as it removes the protecting group, typically a DMT (Dimethoxytrityl) group, from the 5' end of the nucleotide. This activation enables the 5' hydroxyl group of the nucleotide to engage in the next reaction.

Coupling

After deprotection, the activated 5' hydroxyl or amino group is exposed for coupling to the next nucleotide or amino acid. Activators like DCC, HBTU, and DIC are commonly used to enhance coupling efficiency, crucial for synthesis yield and purity.

Oxidation/Condensation

After coupling in nucleic acid synthesis, phosphoramidite bonds must be oxidized to phosphate bonds using oxidizing agents.

Capping

To prevent unreacted ends from becoming by-products in later reactions, a capping reagent (e.g., acetic anhydride) is used to close these ends. This process maintains the purity of the final product by preventing uncoupled precursors from interfering with subsequent synthesis.

After completing the initial four steps, a nucleotide base is linked to the solid phase carrier nucleotide. Acid is then applied to eliminate the protective group DMT from its 5'-hydroxyl group. These steps are reiterated until all necessary synthetic bases are connected.

Highlights of Solid Phase Synthesis Technology for Nucleic Acid Drugs

Our platform utilizes common solid-phase carriers like Controlled Pore Glass (CPG) and polyethylene glycol-based resins (PEG Resin). These carriers can be chosen based on customer requirements to enhance synthesis conditions and enhance the purity and yield of products.
Each cycle of our solid-phase synthesis technology platform includes four key steps: deprotection, coupling, oxidation, and capping. These steps are designed to be efficient and effective, enhancing the quality and efficiency of nucleic acid synthesis.
We provide customized nucleic acid synthesis services to meet specific customer requirements, offering various sequences, lengths, and modifications for research purposes such as gene editing, molecular diagnostics, and drug development.

Custom Nucleic Acid Drug Development Services

CD Formulation offers nucleic acid drug solid phase synthesis technology that is a highly specialized method for manufacturing various types of nucleic acid drugs. These drug types include antisense oligonucleotides (ASOs), small interfering RNAs (siRNAs), messenger RNAs (mRNAs), aptamers, and CpG oligonucleotides.

Custom Small Nucleic Acid Synthesis

Custom ASOs Synthesis
Custom siRNA Synthesis
Custom Aptamer Synthesis
Custom saRNA Synthesis
Custom sgRNA Synthesis
Custom miRNA Synthesis
Custom circRNA Synthesis
Custom shRNA Synthesis
Custom tRNA Synthesis
Custom CpG Oligodeoxynucleotides Synthesis

Custom mRNA Synthesis

Custom mRNA Vaccine Synthesis
Custom mRNA Therapies Synthesis
Custom mRNA Drug Synthesis

Publication Data

Technology: RNA synthesis by solid phase synthesis technology

Journal: Doctoral dissertation, Universität Würzburg

IF: -

Published: 2021

Results:

Deoxyribonucleic acid (DNA) and ribonucleic acid (RNA) are best known for their respective roles in the storage and expression of genetic information. In the last few decades, nucleic acids with a variety of other functions have been discovered or artificially created in living organisms. Examples of these functional nucleic acids include riboswitches, complexes, and nucleases. To obtain information about their functions, a variety of analytical methods can be used. Electron paramagnetic resonance (EPR) spectroscopy is one of several techniques that can be used to study the structure and dynamics of nucleic acids. However, EPR spectroscopy requires unpaired electrons, and nucleic acids themselves are not paramagnetic, so spin labels with free radicals must be added. This paper describes three novel spin labels for analyzing nucleic acids by EPR spectroscopy. They all share two important design features.

Fig.3 Synthesis of RNA in solid phase with phosphoramidites Fig.3 Solid-phase synthesis of RNA with the phosphoramidite method. (Siewert A., 2021)

As a professional provider of solid-phase nucleic acid drug synthesis services, CD Formulation is dedicated to offering high-quality and efficient support for nucleic acid synthesis. Contact us for assistance with your novel nucleic acid drug development and applications.

References

Bege M, Borbás A. The medicinal chemistry of artificial nucleic acids and therapeutic oligonucleotides. Pharmaceuticals, 2022, 15(8): 909.
Siewert A. Nucleotide analogs as rigid spin labels for DNA and RNA. Universität Würzburg, 2021.

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