Protein DNA Conjugation

Protein DNA conjugation, as a bridge, cleverly links the boundaries of protein biochemistry and molecular genetics, and has become the focus of our biopharmaceutical companies in the forefront of scientific research. As a leading CRO, BOC Sciences is focused on providing cutting-edge protein DNA conjugation services that pave the way to scientific innovations and therapeutic breakthroughs.

Protein DNA conjugation technology uses chemical methods to accurately anchor protein molecules to DNA strands, combining the biological potential of proteins with the structural accuracy and programming capabilities of DNA, and is widely permeated in various scientific research and biomedical applications.

Fig. 1 Histone H3-DNA conjugation. (Pujari, S. S., 2021)

Applications of protein DNA conjugation

In the arena of genetic engineering and gene therapy, it has played a key role in the precise regulation of gene expression and the targeted editing of the genome by directing specific proteins, such as transcription factors or nucleases, to DNA sequences. The essence of CRISPR-Cas9 technology is that the Cas9 nuclease fuses with proteins that specifically recognize DNA sequences to ensure precise cutting of the genome. In addition, with the help of the DNA-histone cross-linking strategy mediated by 5-formylcytosine, Pujari et al. were able to construct site-specific complexes and deeply explore the effects of such cross-linking on DNA polymerase activity, further unlocking new mysteries of DNA damage repair and epigenetic regulation.

In the field of cancer therapy, the customized protein-DNA nanoplatform (MnDDC) developed by Nie et al. team provides a novel perspective for collaborative cancer therapy, especially through the Mn²⁺ activated DCV-DNA conjugate, to achieve specific imaging of Mn²⁺ inside the cell, and improve the detection efficiency while ensuring the integrity of the cell.

In the expansion of DNA nanotechnology, the attachment of proteins allows the construction of customized nanostructures, and these complexes show great potential for application in drug delivery, molecular sensing and the development of new materials.

The field of biosensing also benefits from the subtleties of protein-DNA conjugation, which combines high-affinity proteins (such as antibodies) with DNA strands to form highly specific recognition units that can accurately capture target molecules and report their presence through a variety of signaling mechanisms.

In terms of imaging and tracking, visual tags such as fluorescent proteins are incorporated into DNA, which greatly promotes real-time monitoring of the dynamic behavior of DNA in cells, and can be intuitively analyzed at the molecular level, whether it is DNA replication, repair or gene expression process.

Conjugation methods at BOC Sciences

Using amino acid side chain groups: conjugation through amino (-NH2) and sulfhydryl (-SH) groups in proteins is the most direct way. For example, using NHS esters to react with functional groups on a single strand of DNA, covalent bonds can be formed with the amino groups of proteins. On the other hand, maleimide can react with the sulfhydryl group of protein to form a stable covalent connection, especially when the sulfhydryl group of cysteine in the protein acts as the connection point, this way has higher selectivity.

Using special tags: Using genetic engineering techniques to attach special tags to proteins, such as SNAP tags or Halo tags, these tags can specifically bind to the modified DNA sequence, thus achieving covalent conjugation. This method can improve the specificity and controllability of conjugation.

Biotin-streptavidin conjugation: Using the strong biotin-streptavidin interaction, this method provides a robust and high-affinity conjugation pathway that is useful in diagnostic analysis and molecular capture techniques.

Click chemistry: Click chemistry is widely praised for its biological orthogonality and efficiency, allowing it to bind azide-modified DNA to acetylene-modified proteins under mild conditions while maintaining the biological activity of both parts.

Non-covalent interactions: Although less stable than covalent bonding, non-covalent interactions such as aptamer binding to specific proteins and DNA-binding proteins such as zinc finger proteins to DNA sequences are also part of the conjugation strategy. Such methods take advantage of the natural affinity between biomolecules to achieve bonding.

Orthogonal connection strategy: In order to improve the specificity and efficiency of conjugation, multiple connection methods are often combined in research, such as the simultaneous use of linker containing bifunctional groups, such as sulfo-SMCC, to achieve the orthogonal conjugation of different groups.

Protein DNA conjugation services at BOC Sciences

Customized conjugation services: According to the specific requirements of customers, design and synthesize specific sequences of DNA (or oligonucleotide) and select specific proteins, through professional conjugation technology to achieve efficient connection of the two, to meet specific research purposes or application needs, such as the construction of DNA-protein complex for gene editing, disease model construction or biosensing. We also provide oligonucleotides protein oligonucleotide conjugation service.

Chemical modification services: In order to improve conjugation efficiency and product stability, biopharmaceutical companies provide chemical modification services for DNA and proteins, including but not limited to end labeling (e.g., biotin, fluorescent labeling), introduction of reactive groups (e.g., sulfhydryl, amino), and PEGylation to increase water solubility and reduce immunogenicity.

Purification and characterization services: After conjugation is complete, it is essential to provide a high purity DNA-protein conjugate. BOC Sciences usually uses HPLC (high performance liquid chromatography), electrophoresis, mass spectrometry and other technologies for purification, and the conjugate is characterized by enzyme-linked immunosorbent assay (ELISA), SDS-PAGE, dynamic light scattering (DLS) and other methods to ensure product quality.

Functional verification services: In order to verify the biological activity and functionality of the conjugate, we provide a series of biological functional tests, such as intracellular delivery efficiency assessment, transcriptional activation or inhibition assays, enzyme activity assays, and cytotoxicity assays, to help customers evaluate the efficacy of the conjugate in a specific application

FAQ

1. What is protein-DNA conjugation?

Protein-DNA conjugation is a technique that involves chemically linking a protein to a DNA molecule. This hybrid molecule can be used for various applications in diagnostics, therapeutics, and research.

2. Why would I need protein-DNA conjugation services?

Protein-DNA conjugation can be integral for a number of applications, such as targeted drug delivery, development of bioconjugate vaccines, creation of biosensors, and facilitating molecular diagnostics. Our services can provide you with customized conjugates tailored to your specific needs.

3. What types of proteins and DNA can be conjugated?

We can conjugate a wide range of proteins (e.g., antibodies, enzymes) with various types of DNA (e.g., single-stranded, double-stranded, modified oligonucleotides). Our experts can advise on the best combinations and strategies for your specific project.

4. What methods are used for conjugation?

Our conjugation strategies include chemical cross-linking, enzymatic methods, and genetic fusion, among others. We determine the most appropriate method based on the properties of the protein and DNA, as well as the intended application.

5. How do you ensure the stability and functionality of the conjugates?

We employ rigorous quality control processes, including stability testing and functional assays, to ensure that the conjugates maintain their biological activity and structural integrity under specified conditions.

6. What information do you need from me to start a project?

We require details about the protein and DNA sequences, desired conjugation sites or functionalities, the intended application, and any specific requirements or constraints you might have. Providing as much information as possible will help us deliver the best results.

7. How long does the conjugation process take?

The timeline for each project can vary depending on complexity and specific requirements. On average, our standard projects take between 2-6 weeks from initial consultation to final product delivery. We can provide a more precise timeline after assessing your project.

8. Can you help with bulk production?

Yes, we offer scalable solutions for both small-scale and large-scale production. Whether you need milligram amounts for research or gram quantities for industrial applications, we can accommodate your needs.

9. How can I request a quote or place an order?

You can request a quote or place an order by contacting our customer service team via email or phone. Please provide detailed information about your project so we can generate an accurate quote and timeline.

10. Do you offer custom modifications or additional services?

Yes, we offer a variety of custom modifications including labeling, biotinylation, and PEGylation, as well as additional services such as protein expression and purification, DNA synthesis, and analytical characterization. Contact us to discuss your specific needs.