1. Home
  2. Services
  3. Antibody-Oligonucleotide Conjugates (AOC) Development Services
  4. Application of Antibody-Oligonucleotide Conjugates (AOC)
  5. AOC for Proximity Extension Assay (PEA)

AOC for Proximity Extension Assay (PEA)

Antibody-oligonucleotide conjugates (AOCs) represent new technology and have been used in different applications such as imaging, detection, and therapeutics. Creative Biolabs is a recognized leader of high-quality bioconjugation services at an outstanding high success rate. We provide a full set of antibody-oligonucleotide conjugates (AOCs) development services to meet your special needs. The bioconjugates manufactured by us will be stable and effective, which can be successfully applied in various areas such as proximity extension assay (PEA).

Background Our Service Highlights FAQ Published Data

Background

AOC Used for Proximity Extension Assay (PEA) Application

PEA is a direct one-step protein quantification method. It uses a pair of DNA oligonucleotides to connect with the target molecule of the antibody. It needs polyclonal or two monoclonal antibodies (mAbs) to bind closely to the target epitope to form a DNA duplex, which is quantified by real-time PCR.

AOC can be used for the PEA and it is based on the use of two AOC probes, one of which possesses 3′-linked double-stranded DNA, containing a short overhang at its 5′-end. This overhang is used to be complementary to the 5′-linked single-stranded DNA bound to the partner probe. During incubation with mutually recognized samples, hybridization of the overhanging fragments becomes advantageous and the resulting hybridization of double-stranded DNA can be extended by DNA polymerase. This result in a full-length amplicon that can be detected and quantified by qPCR.

Later, this method was optimized so that the double-stranded DNA was replaced by a single strand ON containing complementary sites for direct hybridization with the other ONs, which allowed the extension through the hyper-thermostable DNA polymerase. Furthermore, in multiplex assays, the method can be improved by adding a step of primer digestion after amplification. PEA mainly depends on enzymes, which makes this method relatively expensive and requires special handling and storage. Thus, to overcome this limitation, AOC proximity binding has been combined with polymerization reaction to generate enzyme-free amplification variant of these techniques.

Mechanism of action of PEA probes for protein detection.Fig.1 Mode of action of PEA probes for protein detection. 1

Our Service

AOC for PEA Services

Supported by a team of scientists and experts in the field of antibody development and bioconjugation, Creative Biolabs offers fully integrated AOCs development services for worldwide customers. With extensive experience and advanced technology platform, our conjugation services will be provided in the most high-quality and cost-effective way. We believe our services will help you to make your project a success.

If you are interested in the services and want to know more detailed information, please feel free to contact us and get free consultation and quote. We will get in touch with you as soon as possible and help you design an appropriate protocol to meet your specific requirements.

Highlights

  • Performed by experienced scientists
  • First-class conjugation services with full of technical support
  • Fast turnaround time with professional service
  • Effective and cost-effective

FAQ

  1. Q: What is Proximity Extension Assay (PEA) and how do AOCs contribute to it?

    A: PEA is a sophisticated protein quantification method that utilizes two AOCs. These bioconjugates help form a DNA duplex near the target epitope, enabling precise quantification via real-time PCR, enhancing detection sensitivity and specificity.

  2. Q: How do AOCs enhance the PEA process specifically?

    A: In PEA, AOCs are crucial for bridging two monoclonal antibodies to the target protein. The DNA strands attached to AOCs hybridize, facilitating the extension into a complete amplicon detectable by quantitative PCR, streamlining and amplifying the detection process.

  3. Q: What advancements has Creative Biolabs made in AOC technology for PEA applications?

    A: Creative Biolabs has optimized AOCs for PEA by using single-stranded oligonucleotides that enhance hybridization and enable enzyme-free amplification, making the process more efficient and cost-effective for high-throughput applications.

  4. Q: Can Creative Biolabs customize AOC development for specific PEA requirements?

    A: Yes, Creative Biolabs tailors AOC development to meet unique client needs, offering customized conjugation strategies, oligonucleotide modifications, and assay development to optimize performance in specific PEA setups.

  5. Q: What support does Creative Biolabs offer for researchers using AOCs in PEA?

    A: Creative Biolabs provides comprehensive support for AOC-PEA applications, including expert consultation, protocol design, and fast, high-quality service to ensure effective results. Researchers can access customized solutions and technical assistance to seamlessly integrate AOCs into their diagnostic workflows.

Published Data

The Proximity Extension Assay (PEA) played a crucial role in this experiment by enabling the sensitive detection and quantification of proteins. In the study, antibody-DNA conjugates were created, which involved conjugating specific antibodies with DNA oligonucleotides. These conjugates were then used in a proximity ligation assay, where two antibody-DNA conjugates that recognize different epitopes of the target protein (HIV p24) were brought into proximity, facilitating the formation of amplifiable DNA sequences upon ligation. The assay was performed in a solid-phase format, with biotinylated capture probes and streptavidin-coated magnetic beads used for capturing and isolating the complexes. This method allowed for highly sensitive detection of the target protein, with detection limits between 10 and 100 pg/ml, demonstrating the assay's robustness and sensitivity in quantifying protein concentrations in complex biological samples.

Proximity ligation assay for HIV p24 using constructed AOC.Fig.2 Proximity ligation assay of HIV p24 using AOCs from tagged derivatives of clone 1 and and clone 2.2

References

  1. Dovgan, Igor, et al. "Antibody–oligonucleotide conjugates as therapeutic, imaging, and detection agents." Bioconjugate Chemistry 30.10 (2019): 2483-2501.
  2. Soxpollard, Noah, et al. "Selection of antibody-binding covalent aptamers." bioRxiv (2023): 2023-03.

For Research Use Only. NOT FOR CLINICAL USE.

Services
ADC
Related Sections
Antibody-Oligonucleotide Conjugates (AOC) Development Services:
Services:

Online Inquiry

Name:
*Phone:
*E-mail Address:
*Products or Services Interested:
Company/Institution
Project Description:

Welcome! For price inquiries, please feel free to contact us through the form on the left side. We will get back to you as soon as possible.