Oligonucleotide Services

Highly Customizable and Cost-Effective Oligonucleotide Services

NJ Bio offers innovative solutions for oligonucleotide synthesis within highly standardized process controls. Our extensive technical expertise enables us to provide a broad range of chemistries and modifications in oligonucleotide sequences. We can also deliver oligonucleotide conjugates for a variety of applications and constructs.

At NJ Bio, we synthesize unmodified and modified oligonucleotides with defined sequences using Phosphoramidite solid phase synthesis. These oligonucleotides are then purified by HPLC (RP and/or IEX) to achieve the desired purity. Using our state-of-the-art facility for oligonucleotide synthesis, we can accelerate your drug discovery program.

Why should you choose NJ Bio for your oligonucleotide research needs?

NJ Bio can tailor the process of oligonucleotide synthesis with respect to the required length, sequence and modifications based on your research needs. We offer various chemical modifications including phosphorothioate linkages, conjugation with biomolecules and other functional groups that are added to enhance stability, specificity, or detection of the oligonucleotide. Our experience in bioconjugation and small molecule therapeutics enables us to devise the best approach to help you design your oligonucleotide-based therapeutic with the best efficiency. Our research expertise also enables us to optimize the oligonucleotide molecules for better bioavailability and function. We utilize robust analytical instruments and optimum procedures that are amenable to synthesis of oligonucleotides with high accuracy and uniformity to give our clients the best experimental results.

NJ Bio stays up to date with the latest advancements in oligonucleotide synthesis technology. Our state-of-the-art facility is equipped with an automated Oligo synthesizer and advanced purification systems which allow us to obtain high yields, purity, and quality. We understand that each project is unique, and we work closely with our clients to design custom oligonucleotides that meet their specific needs. Our rigorous quality control measures include thorough purification, analytical testing, and documentation of the synthesis process. The streamlined processes and efficient workflows allow us to provide fast turnaround times, without compromising on the quality of the products. Our team is responsive, knowledgeable, and dedicated to assisting you through the entire oligonucleotide synthesis process, from project consultation to post-synthesis support.

Solid-Phase Oligonucleotide Synthesis

At NJ Bio, we pride ourselves on providing the highest quality of service and material to our clients. We recently invested in a state-of-the art automated Oligo Synthesizer, the Ӓkta OligosyntTM, which allows us to carry out custom synthesis of unmodified and modified oligonucleotides with defined sequences. The Ӓkta OligosyntTM oligo synthesizer covers a wide scale range (0.2 to 12 mmol); this system is best suited for sequences ≤100 base pairs while sequences ˃100 base pairs are more reliably prepared using PCR, in vitro transcription, and cell-based methods. Though these processes limit flexibility, they offer the advantage of synthesizing longer nucleotide sequences. At NJ Bio, we help you choose the most suitable approach for your program to obtain high yield with best sequence fidelity and least errors.

The process of custom synthesis generally involves several steps, including the design and optimization of synthetic routes, chemical synthesis and purification, characterization and analysis of the synthesized compounds, and quality control to ensure the purity and quality of the final product. NJ Bio can customize this process accordingly to cater to the customers research needs.

Fig. 1: Process of Custom Synthesis of Oligonucleotides

Fig. 1: Process of Custom Synthesis of Oligonucleotides

Case Study 1:

At NJ Bio, we have synthesized a variety of different oligonucleotides containing numerous modifications. These include standard DNA and RNA oligos, siRNA and ASOs; most notably the synthesis of a 5-10-5 gapmer ASO for further conjugation with a synthetic peptide. The 5-10-5 gapmer oligonucleotides are the second-generation of antisense molecules; their design incorporates two “wings” of 5 bases (modified with a nuclease-resistant/enhanced-binding-affinity sugar modification)¹ and a central gap of 10 standard deoxyribonucleotides (no sugar modification) that allow for RNase H binding. We were able to successfully synthesize multi-gram (~5g) quantities of a 5-10-5 gapmer at high purity (>90%) using our Ӓkta OligosyntTM oligo synthesizer. The synthesized ASO was further modified during the synthesis cycle to incorporate an amine-functionalized linker that allowed for additional modification and conjugation with a biomolecule. The final conjugate was isolated by prep-HPLC and desalted using SEC.

Fig. 2. Ӓkta Oligosynt^TM oligo synthesizer, NJ Bio

Case Study 2:

Using our wealth of chemistry and bioconjugation experience, we can employ a wide variety of post-synthetic modifications. A major post-synthetic modification includes addition of a bifunctional cross-linker which enables us to conjugate the synthetic oligonucleotides with larger biomolecules such as monoclonal antibodies (mAbs), carbohydrates, lipids as well as peptides and proteins. Additionally, we have experience in hybridizing complementary oligonucleotide strands (siRNA) as well as carrying out enzymatic ligation of oligonucleotides containing 5′ and 3′ phosphate groups (aptamers).  NJ Bio was able to fully optimize the enzymatic ligation of two oligonucleotide strands to form an aptamer that was further modified to allow for conjugation with mAb. The aptamer was extensively characterized at every step using HPLC (RP-IP) as well as LC-MS (ESI) to ensure minimal by-product formation. The aptamer was then functionalized with a PEGylated linker and subsequently site-specifically conjugated with an antibody. The desired conjugate was isolated using SEC and characterized on SDS-PAGE.

Fig. 3. LC-MS unit at NJ Bio

Case Study 3:

NJ Bio is not only capable of preparing small amounts of oligonucleotides-conjugates (1-50mg) but also has successfully prepared up to 50g of oligonucleotide conjugates. With equipment such as the Ӓkta Pilot chromatography system, we can purify multiple grams of material in a single run, which expedites the overall process of synthesis. For this, we prepared a large-scale oligonucleotide-peptide conjugate using a traditional maleimide conjugation strategy. Following the conjugation of the oligonucleotide (siRNA) with a protein, we induced ring-opening of the unstable Thiosuccinimide to generate the more stable thioether². Following ring-opening the material was purified twice to remove any excess oligonucleotide (IEX) and unconjugated protein (HIC). The identity and purity of isolated conjugate was confirmed by a variety of analytical techniques including LC-MS, SEC, and SDS-PAGE.

Fig. 4. Purification Unit

Analytical Support

Analytical characterization plays an important role in ensuring the product’s purity and integrity for safety and efficacy purposes. However, the unique charge and complex structure of oligonucleotides makes the analytical characterization challenging. Selection of right analytical tools depending on the molecule size, higher order structure and chemical modifications is crucial to obtain the best results. We use some of the best analytical instruments and methods with high resolution and sensitivity such as QqQ, TOF & Q-TOF Mass Spectrometers, advanced chromatographs, electrophoresis systems, and other plate-based assays to analyze the physicochemical and biophysical properties of oligonucleotides, ensuring the final product is of high quality.

NJ Bio offers the following Oligonucleotide Services:

• Automated DNA/ RNA oligonucleotide synthesis
• All standard modifications with or without backbone modifications
• Oligo Bioconjugation
• Nucleic acid analogs such as Bridged Nucleic Acid (BNA), Locked nucleic acid (LNA), etc.
• Analytical services (Method Development, Method Validations/Qualifications and Transfers)
• Quality Control and Stability analysis
• Regulatory support
• Multigram oligonucleotide synthesis and purification for pre-clinical market