The Importance of Oligonucleotide Purification

Synthetic oligonucleotides are in increasing demand in fields ranging from clinical diagnostics to novel biopharmaceutical therapeutics. The synthesis of oligonucleotides is an efficient process, and small amounts of impurities are produced during each step of the synthesis. Therefore, manufacturing organizations as well as individuals who rely on the quality of oligonucleotides have shown interest in developing effective and efficient methods for the purification and analysis of these important biological aptamers. For researchers, there is also a need to use oligonucleotides of higher purity. Failure to purify crude synthetic oligonucleotides can seriously hamper an organization's or individual's ability to achieve desired results. Therefore, oligonucleotides used for gene knockout, genotyping, and diagnostic purposes often require further purification after synthesis.

Key Technologies

Our purification service is built upon a foundation of established and advanced purification techniques. While a simple desalting step effectively removes small molecules and salts, it is not sufficient to eliminate failure sequences and other product-related impurities. Therefore, our service integrates specialized chromatographic methods to ensure comprehensive purification. This multi-faceted approach allows us to not only remove residual synthesis reagents and salts but also to separate the full-length target oligonucleotide from shorter, incomplete fragments, which are a major byproduct of solid-phase synthesis. Our expertise in tailoring these techniques to the specific properties of each oligonucleotide fragment guarantees a product that is pure, stable, and ready for use in a wide range of demanding applications.

Desalt to Perfect: Precision-Purified Oligos.

CD BioGlyco has a professional Glyco™ Synthesis Platform with multiple world-leading technologies to provide clients with outstanding Custom Carbohydrate Synthesis. CD Bioglyco provides technologies based on C-18 reversed-phase high-performance liquid chromatography, technologies based on normal-phase chromatography, and technologies based on gel filtration to desalt oligonucleotides. Desalting is a very basic oligosaccharide purification step. In this step, excess salt is removed from the mixture by using normal-phase chromatography. It will not eliminate the fault sequence. It produces salt-free, ready-to-use DNA solutions ideal for robust techniques such as polymerase chain reaction (PCR), amplified restriction fragment polymorphism analysis, microarrays, and more. Desalting also allows for use in combination with other purification techniques.

Additionally, we offer chemical-based methods for desalting oligonucleotides. For example, oligonucleotides are desalted by ethanol precipitation. Briefly, an equal volume of water is added to a solution of triethylamine trihydrofluoride, which is then diluted with slightly acidic sodium acetate. After adding 100% ethanol for precipitation, followed by high-speed centrifugation.

Fig.1 Desalination-based oligonucleotide fragment purification technology. (CD BioGlyco)

Workflow

• Initial Desalination

This crucial first step removes excess salts and small-molecule impurities that are byproducts of the synthesis and deprotection process. This initial cleanup is essential for preparing the sample for subsequent, more sophisticated purification steps.

• Target Fractionation

The desalted sample is then subjected to a high-resolution chromatographic step. This stage is designed to separate the full-length oligonucleotide from the shorter failure sequences based on their physical properties, such as size and charge. This is the key step where product purity is significantly enhanced.

• Final Desalination & Concentration

The highly purified fraction containing the target oligonucleotide is collected. A final desalting step is performed to remove any remaining buffer salts from the chromatographic process. The final product is then concentrated and dried, ready for shipment and downstream use.

Publication Data

Applications

• Desalting technology is an important processing step before oligonucleotides are subjected to PCR.
• Desalting technology is an important processing step before oligonucleotides are subjected to amplified restriction fragment polymorphism analysis.
• Desalting technology is an important processing step for oligonucleotides before microarray analysis.

Advantages

• By combining state-of-the-art chromatography with advanced desalting techniques, we deliver oligonucleotides with superior purity while minimizing product loss. Our published data demonstrates an improved yield compared to standard purification methods.
• Our optimized workflow and sophisticated equipment allow for efficient processing, ensuring you receive your purified oligonucleotides promptly to keep your research on schedule.
• Every oligonucleotide fragment we purify undergoes a rigorous quality control process, including mass spectrometry and analytical HPLC, to guarantee its identity and purity.

Frequently Asked Questions

Associated Services

Desalination purification efficiently removes salts and small impurities via precipitation or filtration, providing a cost-effective solution for basic applications like short oligos where truncated sequences are tolerable. However, for applications demanding near-absolute purity is essential to eliminate critical contaminants. To achieve this, we offer specialized associated services:

CD BioGlyco is committed to providing the best solutions for Custom Oligonucleotide Synthesis for global clients. We provide high-quality Oligonucleotide Fragment Purification Services for clients all over the world. If you are interested in our services, please feel free to contact us for more details.