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Oligonucleotide Synthesis

Oligonucleotide Synthesis

Aurigene is one of the leading service provider in oligonucleotide synthesis with capabilities to perform oligo synthesis, characterization and biological evaluation.

The oligonucleotide synthesis services at Aurigene is supported by a state-of-the-art lab facility and skilled scientists with expertise in various types of oligos. We have an in-house custom synthesis of phosphoramidites and the ability to perform complex conjugation using click chemistry, amide coupling and thiol mediated C-S & S-S bond.

Our dedicated lab space of 1000 sq.ft is equipped with humidity control. Aurigene’s oligonucleotide platform is supported by trained team that helps optimize the designed oligonucleotide along with primary assay iterations. We have the ability to synthesize 10-40-mer oligonucleotide up to 1-gram scale. We perform all standard modifications, with or without backbone modification, for which phosphoramidites are made in-house and also available commercially. Oligo equipments at Aurigene: K and A H-16 automated DNA/RNA synthesizer AKTA pure 25M FPLC purification system Sartoflow smart TFF system for concentration and desalting Martin Christ LSC plus lyophilizer with lyocube, benchtop tray lyophilizer Agilent LC-MS for oligonucleotide analysis Implen N60 nano spectrophotometer

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Synthesis and Purification

  • Scale: 1-100 µmol (1-100 mg)
  • 16x1 µmol, 4x20 µmol, 1x100 µmol
  • Size: 10 to 40-mer
  • 2’-F, 2’-OMe, 2’-O-MOE, LNA and all standard modifications
  • Anion/ Cation exchange purification
  • Reverse phase HPLC
  • Size exclusion
  • Desalting

Analysis

  • UV quantification
  • Purity and identity analysis using LC-MS (TOF) and mass deconvolution
  • HPLC with different mobile and stationary phase for purity assessment

Biology

  • In vitro IC50, Kd and binding assessment and in vivo activities evaluation with wide range of cells. Efficacy experiment in oncology and inflammation model
  • Toxicology studies with ASO.

Conjugation

  • Acid-amine coupling
  • Alkyne-azide coupling
  • Thiol-maleimide coupling
  • S-S and C-S bond formation

Why Aurigene Pharmaceutical Services?

Complex conjugation using click chemistry, amide coupling and thiol mediated C-S & S-S bond

Ion exchange and reverse phase HPLC

Expertise in nucleoside, sugar modifications, base modifications

State-of-the-art drug discovery facility using oligonucleotide modality

CADD Optimization

Synthesis between 10-40-mer oligonucleotide

In-house custom phosphoramidite synthesis

Integrated Services

Virtual Tour

 

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Frequently asked questions

How are oligonucleotides synthesized?

Oligonucleotides can be synthesized using solid-phase (Peptides) and one pot liquid phase (enzymatic) method in the lab for milligram to multi-kilogram scale. Steps involved in solid-phase Oligo synthesis are detritylation, coupling, oxidation and capping. One-pot liquid phase oligo synthesis requires polymerase enzyme, template oligonucleotide and nucleotide triphosphate as starting materials.

How accuracy of chemical synthesis of oligonucleotides is maintained?

Accuracy of chemical synthesis of Oligonucleotides is maintained by maintaining the order of starting materials used during the synthesis process.

What are oligonucleotide drugs?

Oligonucleotide drugs help modulate the effect of a disease-causing gene by targeted degradation/activation of gene expression or gene editing. This activity may cause the disease to be controlled or cured.

How stable are the Oligos?

Oligos are stable for 6 weeks in buffer when stored at 37°C and for long-term stability is to be stored between -20°C to -80°C. During storage, oligos should not be exposed to direct sunlight or UV rays.

What is the oligo synthesis scale?

We can Synthesis 10-40-mer oligonucleotide up to 1 gram scale.

Can you incorporate a specific modification into Oligo?

We can perform all standard modifications, with or without backbone modification, for which Phosphoramidites are commercially available. We also have the capabilities of making any customized Phosphoramidite in-house.

What are the recommended storage conditions for Oligonucleotides?

For a period of 6 weeks, Oligos can be stored in T10E1 buffer at 37°C and for the long term, Oligonucleotides can be dried down and stored with or without TE buffer at -20°C.

Does Oligonucleotide have a phosphate on the 5’ or 3’ end?

Typical Oligonucleotide synthesis does not produce phosphate on the 5’ or 3’ end. On request, oligonucleotides can be synthesized with phosphate on the 5’ or 3’ end.

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