Oligonucleotide Bioconjugation

Service Description

The conjugation of oligonucleotides with various molecules plays a pivotal role in genetic research, drug development, and diagnostics. As a professional biotech company, BOC Sciences offers custom bioconjugation services. We provide top-notch services related to oligonucleotide conjugation, supporting oligonucleotide modifications at specific sites, and helping customers' oligonucleotide therapeutics development.

What are Oligonucleotides?

Oligonucleotides are short DNA or RNA molecules that have a wide range of applications in genetic testing, research, and forensics. They are typically composed of 13 to 25 nucleotides; this small size allows for precise binding to specific sequences in the DNA or RNA of an organism, making them useful in techniques such as PCR, molecular cloning, and DNA sequencing. The four types of nitrogenous bases in DNA oligonucleotides are adenine (A), thymine (T), guanine (G), and cytosine (C), while RNA oligonucleotides include uracil (U) instead of thymine. The specific sequence of these bases in an oligonucleotide determines its function and interaction with other molecules.

Fig. 1 Oligonucleotide conjugates

Methods of Oligonucleotide Conjugation

BOC Sciences will employ the appropriate conjugation methods according to the customer's needs to attach the desired molecule to the oligonucleotide without disrupting its structure and function.

Chemical Conjugation: Common reactive groups used for chemical conjugation include -NH₂, -SH, and -COOH.

Enzymatic Ligation: Common enzymes used for oligonucleotide conjugation include terminal transferases, ligases, and transglutaminases.

Solid-Phase Synthesis: It is a powerful technique used in the efficient and automated production of oligonucleotides, allowing the attachment of molecules to oligonucleotides in a controlled manner.

Click Chemistry: Commonly used click chemistry reaction for oligonucleotide conjugation include CuAAC, SPAAC.

Photocrosslinking: The photoreactive groups are introduced to the oligonucleotide and the molecule to be attached.

Biotin-Streptavidin Based Conjugation: Biotin-streptavidin-based conjugation relies on the strong and specific interaction between biotin and streptavidin.

Oligonucleotide Conjugates Purification

After conjugation, the purification of oligonucleotide conjugates becomes crucial to ensure their functionality. We use various purification techniques to remove impurities, unreacted molecules, and incomplete conjugates.

Oligonucleotide Conjugates Analysis

We utilize several analytical techniques to characterize these conjugates, providing valuable insights into their structure, purity, stability, and interactions. Moreover, we can provide complete analysis reports and experimental data.

Advantages of Oligonucleotide Conjugation

High Specificity: Oligonucleotides, such as DNA, RNA, and PNA (peptide nucleic acid), can easily bind to their complementary strands with high specificity, allowing for precise targeting in biological applications.

Versatility: Oligonucleotides can be conjugated with various other compounds, including proteins, peptides, fluorescent dyes, and nanoparticles, providing a great deal of versatility in their use.

Structure Visualization: The attachment of fluorescent dyes or other labels to oligonucleotides allows for the visualization of molecular structures and processes in real time, aiding in the understanding of biological systems.

Enhanced Stability: Conjugation can enhance the stability of oligonucleotides in biological systems, increasing their resistance to degradation by nucleases.

Delivery Efficiency: Conjugation with specific targeting ligands or nanomaterials can improve the delivery efficiency of oligonucleotides to target cells or tissues.

Applications of Oligonucleotide Conjugation

• Targeted Drug Delivery

By coupling therapeutic agents with oligonucleotides, researchers can achieve highly specific drug delivery to specific cells or tissues, minimizing off-target effects and reducing systemic toxicity. Additionally, the ability to modify oligonucleotides allows for the fine-tuning of drug release kinetics.

• Therapeutic Oligonucleotides

Modified oligonucleotides can be designed to target disease-causing genetic mutations, inhibit the expression of harmful proteins, or correct genetic defects.

• Diagnostic Assays

Oligonucleotide conjugates have been instrumental in enhancing the sensitivity and specificity of diagnostic assays. By coupling oligonucleotides with fluorescent or radioactive labels, researchers can create highly sensitive probes for detecting specific DNA or RNA sequences. Furthermore, the modified oligonucleotides have been served as ideal tools for a range of diagnostic applications, including PCR-based assays, in situ hybridization, and nucleic acid microarrays.

• Therapeutic Oligonucleotides and Nanotechnology

Oligonucleotides can be attached to various nanoparticles, such as liposomes, polymeric nanoparticles, or gold nanoparticles, enabling targeted delivery and controlled release of therapeutic oligonucleotides. These nanoscale delivery systems offer unique advantages, such as prolonged circulation time, enhanced cellular uptake, and reduced immunogenicity.

• Immunotherapy

Oligonucleotide bioconjugates can function as immune stimulants or antisense molecules, making them valuable tools in the development of new immunotherapies.

• Research Tools

In lab research, bioconjugated oligonucleotides can be used for various purposes, such as studying gene expression, investigating protein-DNA interactions, or tracking the location and movement of specific molecules within cells.

Our Oligonucleotide Conjugation Services

Custom Synthesis: We provide customized synthesis of oligonucleotides to meet unique needs of our clients, fulfilling the exact sequence, length, and structure as required for specific applications.

Oligonucleotide Modifications: We offer a variety of modifications including fluorescent and non-fluorescent labeling, ligation with proteins or peptides, end-capping, phosphorothioate modifications, biotinylation, and more.

• Antisense oligonucleotide (ASO) conjugation

Antisense oligonucleotides are short, synthetic nucleic acid strands that can bind to specific messenger RNA (mRNA) sequences to inhibit gene expression. ASOs can be conjugated to various molecules to improve their stability, cellular uptake, or targeting capabilities. Common conjugation strategies include linking ASOs to: peptides or proteins, lipids and polyethylene glycol (PEG).

• siRNA conjugation

Similar to ASOs, siRNA molecules are designed to silence specific genes by targeting mRNA. Conjugation of siRNA can enhance their delivery and efficacy. Common conjugation approaches for siRNA include: lipid-based formulations, polymer-based conjugates, antibodies and peptides.

• microRNA conjugation

MicroRNAs (miRNAs) are small non-coding RNAs that regulate gene expression. Conjugation strategies for microRNAs are similar to those for siRNA and ASOs, focusing on enhancing delivery and stability. Some common conjugation methods include: lipid-based delivery systems, polymer conjugates and exosome-based delivery.

• Aptamer conjugation

Aptamers are short, single-stranded DNA or RNA molecules that can bind to specific targets with high affinity and specificity. Conjugation of aptamers can be used to target therapeutic agents or imaging probes to specific cells or tissues. Common methods of aptamer conjugation include: fluorescent dyes or imaging agents, drug conjugates and nanoparticles.

Analytical Testing and Validation: Our services include comprehensive testing and validation to ensure the integrity and efficacy of oligonucleotides. This includes purity analysis, quantification, and structural verification.

Large Scale Production: We are equipped to handle large-scale production of oligonucleotides, ensuring consistent, high-quality products to cater to both industrial and research needs.

Troubleshooting and Optimization: Our team of experienced scientists provide troubleshooting and optimization services, addressing any issues with the synthesis or application of oligonucleotides, and improving their overall performance.

Consultation: We provide expert guidance and consultation on project design, experiment setup, result interpretation, and other related aspects.

Our Competitive Advantages

Scientific Expertise: Our team of scientists has years of experience in the field of Oligonucleotide Bioconjugation, enabling us to provide high-quality services in this specialized area.

Advanced Technology: We use state-of-the-art technology and techniques that allow us to be accurate and efficient in our service delivery.

24h-Available Customer Services: We offer personalized solutions to our clients to meet their unique requirements in Oligonucleotide Bioconjugation.

Quality Control: We adhere to strict quality control procedures to ensure we deliver reliable and credible services to our clients.

Fast Turnaround Time: Our refined processes and experienced team allow us to deliver services in a timely manner without compromising on quality.

Excellent Customer Service: We uphold the highest standards in customer service to ensure complete client satisfaction.

Competitive Pricing: We provide our top-notch services at competitive prices, ensuring value for money.

Confidentiality: We respect our client's proprietary information and ensure the utmost confidentiality of any project-related data.

Constant Innovation: We are dedicated to continuous research and development to stay ahead of advancements in the field and provide the latest solutions.

Global Reach: We offer our services globally, being able to cater to clients across different geographic locations and time zones.

Quality Services

Oligonucleotide Conjugation

Oligonucleotide bioconjugation refers to the process of chemically attaching oligonucleotides to other substances such as proteins, peptides, or other oligonucleotides. The resulting oligonucleotide conjugates leverage the strengths of both the oligonucleotide and the attached molecule, unlocking potential for targeted interactions within biological systems. By linking oligonucleotides with various molecules like peptides, antibodies, fluorophores, or drug molecules, BOC Sciences can create oligonucleotide conjugates with unique properties and functions.

Case Study

• Case Study 1

Gene therapy has offered promise for treating gene-related diseases. Synthesis and modification simplicity, physiological safety and stability are some of the merits that have contributed to the success of LONs in the delivery of gene molecules for the induction or knockdown of specific gene expression. This research reported two methods for conjugating specific RNA strands with hydrophobic chains, allowing the formation of RNA-amphiphiles and subsequent efficient gene-silencing capability. By employing dibenzocyclooctyne group (DBCO)-labeled DNA-amphiphile as a template, azide-modified RNA was directly attached to the hydrophobic chain via 'click-chemistry' and the hybridized DNA could then be removed by adding DNase I. This interesting method provided a novel solution to the challenge of linking RNA strands with hydrophobic molecules, and the outcome has improved the efficacy of RNA therapeutics.

Fig.2 Gene-silencing using RNA amphiphiles. (Dore, M. D., 2018)

• Case Study 2

In this study, lipid nanoparticles (LNPs) loaded with siRNA targeting Polo-Like Kinase 1 (PLK1) protein, present in the triple negative breast cancer cell line (MDA-MB-231), have been modified with antibodies to target tumors. Biodistribution studies of labeled siRNA-LNPs demonstrated that antibody modified LNP (antibody against heparin-binding EGF-like growth factor, αHB-EGF) effectively delivered siRNA to tumor tissue in mice. Interestingly, the PLK1 protein expression was inhibited after intravenous injection of the LNPs and tumor growth was decreased. These results indicate that antibody modified LNPs loaded with siRNA are a promising therapeutic approach for breast cancer.

Fig.3 Anticancer effect of αHB-EGF LNP-siPLK1 in vivo. (T Okamoto, A., 2018)

Resources

• siRNA Delivery Optimization and Development

FAQ

1. What types of linkers do you use in the oligonucleotide bioconjugations?

We use several types of linkers in our oligonucleotide bioconjugations. These include disulfide linkers, thioether linkers, and maleimide linkers among others.

2. Can I supply my own oligonucleotide for bioconjugation?

Yes, you may certainly provide your own oligonucleotide for bioconjugation. However, we will first conduct a thorough evaluation to ensure that it meets our quality standards.

3. How long does the bioconjugation process usually take?

The time frame can vary depending on the complexity of the project. Generally, it takes around 2-3 weeks. We will provide an estimated timeline after we carefully evaluate the project requirements.

4. Will I retain intellectual property rights of the bioconjugated molecule?

Yes, any service project we undertake is strictly confidential, and you will retain all intellectual property rights related to your bioconjugated molecule.

5. Can I order a custom bioconjugation service?

Absolutely. We offer custom bioconjugation services tailored to the specific needs of your research project or application.

6. How do you handle the stability and storage of conjugated oligonucleotides?

Your conjugated oligonucleotides will be delivered lyophilized, protected from light, and under conditions that ensure the maintenance of product stability. We recommend storing the product at -20 °C for long-term use.