Synoligo boosts production with new facility, expanding capacity for oligonucleotide synthesis. Read the press release

Your Blueprints,
Our Expertise

Start Your Experiment With High Quality Oligos​.

Stages of Synthesis

Purification Methods

Purification Methods

01
Desalting
This method can remove salts and residual solvents but not truncated or incomplete sequences
02
Gel Electrophoresis
PAGE Gel is often used to separate oligos based on molecular size. Full length products are isolated by running them through a polyacrylamide gel matrix.
03
High-Performance Liquid Chromatography (HPLC)
Different types of HPLC can be used to separate full length product from impurities.

Reverse Phase (RP-HPLC)

Commonly used to check for purity of modified or labeled oligos. This method separates based on hydrophobicity.

Anion Exchange (AEX-HPLC)

Commonly used for phosphorothioate-modified oligos and it is separated by charge.

Hydrophobic interaction chromatography (HIC)

Hydrophobic interaction chromatography separate molecules based on their surface hydrophobicity. Contrary to AEX, product elutes in low salt concentration. Many applications have been published using this technique for DMT-ON purification and then on-column DMT cleavage

Post Purification QC Methods

Molecular Weight by Mass Spectrometry
The most preferred method for diagnostic and therapeutic oligos. It is used to confirm the integrity and the molecular weight of the oligo. In ideal scenarios, oligo sequence information can be inferred from MW determination. To get absolute confirmation of sequence, MS/MS or NGS has been used.
Purity by Chromatography
Liquid chromatography is commonly used to assess the purity of the full-length product and gauge product quality. Separation can be based on reverse-phase, ion exchange, size exclusion, or hydrophobic interaction. In high-throughput modes, a generic LC method (typically reverse-phase) is often used, prioritizing speed over quality. However, in therapeutic development, even minor changes in sequence can lead to vastly different peak resolutions, meaning the generic method often overestimates product purity and doesn't differentiate all impurities from the full-length product. For clinical applications, LC methods are typically sequence-specific, and two or more orthogonal methods are often used to fully characterize the product, as no single method resolves all impurities.
Moisture Content
Oligonucleotides are highly hygroscopic, meaning they readily absorb moisture. No matter how long you dry the product, residual water will always remain. This can be determined following USP <921> guidelines.
Sodium Content
High sodium content can disrupt ion pairing during ion-pair reversed-phase liquid chromatography (RP-LC) analytical separation, altering retention times and causing peak splitting. Additionally, sodium ions can interfere with downstream quality control processes, particularly in mass spectrometry, by suppressing signals due to their high ionization energy and competing with other analytes for ionization. Therefore, measuring and desalting are essential steps to ensure high-quality control of oligonucleotides.
Endotoxin
Testing ensures that the toxic component, Lipid A, is at or below levels recommended by the United States Pharmacopeia (USP) and FDA. Read our blog post to learn more about the various tests used to assess safety levels.
Heavy Metal
Screening for metal contamination is becoming increasingly important for product safety. ICH classifies heavy metals into three classes, 1, 2, and 3. Class 1 metals, including As, Cd, Pb, Hg, should be essentially absent as they are known or strongly suspected human toxiants. Class 2 metals, divided into two subcategories A and B based on their probability of occurring in the drug product, include Co, Ni, V as A and Ag, Au, Ir, Os, Pd, Pt, Rh, Ru, Se, and Tl. Class 3 metals has low toxicities in general including Ba, Cr, Cu, Li, Mo, Sb, and Sn.
Residual Solvents
Residual solvents are categorized into three classes based on toxicity, with Class 1 being the most toxic. To meet high product safety requirements, testing for residual solvents used during synthesis, purification, or in excipients or drug products is necessary. Customers can choose to test the final product or each individual component.
Bioburden
This test is crucial for determining the number of microbes present in the product. Microbial extraction methods include mechanical testing, vortexing, and sonication. The extract fluid is then used for culturing different organisms and tested by pour plating or filtration.

Synoligo's Competitive Advantage

State-Of-The-Art Equipment​

Optimal efficiency in production, purification and lyophilization

Large Selection Of Raw Materials for Common and Rare Modifications​

Enable quick start of your project​

Specialization in Complex Oligo Modifications

Can take on custom projects that are difficult or do not fit in standard workflow from other CROs​

Automated Workflow​

Can meet any demands for low & high throughput (column & plate formats)

Stringent QC Method​

Can guarantee accuracy, yields and quality based on the project​

Additional Services with Custom Project

Process transfer for final product manufacturing – clinical use

Popular Modification

Synoligo synthesized oligos with common modifications daily. Our specialization is manufacturing oligos with complex modifications. ​Below is a list of popular modifications that can be synthesized for DNA/RNA oligos. Contact us for more information on modifications that fit your need.​

Common Modifications
Attachment Modifications
Biotin
BiotinTEG
Thiol-C6 S-S
Aminolinker
Thiol-C3 S-S
dTAminolinker
Quencher 
MGB
Eclipse
BHQ1/2
and more
Dabcyl
Fluorescent Dye
ROX
TAMRA
CY5
FAM
TET
CY3
HEX
Cy5.5
and more
Modified Bases
dI
dU
Phosphorothioate
and more
Other Custom Mods
Peptide
Enzymatic Ligation
Protein
5' and 3' Phosphorylation (Phosphate)
Antibody
TNA
BNA
MOE
FANA
GNA

Applications of Oligos

With decades of experience, our expert team specializes in manufacturing oligos tailored to diverse scientific applications, ensuring your specific needs are met.

Research & Innovation

Extensive list of customization allows researchers to achieve their research goals.

Molecular Diagnostics

Broad selection of various dyes and probes.

Therapeutic Development

Guarantee ultra low endotoxin even for lipid modified oligo.

research-icon
molecular-icon
therapeutic-icon
  • Branched/dendrimer
  • Circular DNA/RNA
  • Ultra-long oligos (150-200nt)
  • DNA-encoded library (DEL)
  • Primers PCR/qPCR
  • FISH/MERFISH Probes
  • FRET Probes
  • NGS Adapters
  • Molecular Beacon
  • Antisense
  • Immunostimulatory
  • mi-/sa-/siRNA​
  • Aptamer
  • CRISPR gRNA​

Applications of Oligos

With decades of experience, our expert team specializes in manufacturing oligos tailored to diverse scientific applications, ensuring your specific needs are met.

Research & Innovation

Extensive list of customization allows researchers to achieve their research goals.

research-icon
  • Branched/dendrimer
  • Circular DNA/RNA
  • Ultra-long oligos (150-200nt)
  • DNA-encoded library (DEL)

Molecular Diagnostics

Broad selection of various dyes and probes.

molecular-icon
  • Primers PCR/qPCR
  • FISH/MERFISH Probes
  • FRET Probes
  • NGS Adapters
  • Molecular Beacon

Therapeutic Development

Guarantee ultra low endotoxin even for lipid modified oligo.

therapeutic-icon
  • Antisense
  • Immunostimulatory
  • mi-/sa-/siRNA​
  • Aptamer
  • CRISPR gRNA​
To top