Diels-Alder Reaction

Bodies contain many different molecules, such as proteins, lipids, glycans, and small molecules, and they have many functional groups, including alcohols, amines, carboxylic acids, and thiols. These reactions can proceed in a living and are referred to as bioorthogonal. The Diels–Alder reaction is a chemical reaction of dienophiles that form a substituted cyclohexene derivative, which is a pericyclic reaction with a concerted mechanism. The diels–alder reaction provides a reliable way to form six-membered rings by the simultaneous construction of two new carbon-carbon bonds, which can be good control over the region and stereochemical outcomes.

Fig.1 The diels–alder reaction. (Wang, et al., 2023)

Diels-Alder Reaction-based Cell Surface Chemical Conjugation Glycoengineering Service at CD BioGlyco

CD BioGlyco possesses the most advanced equipment and unique R&D expertise to provide personalized according to client's specific needs promptly. The chemical conjugation service offered by CD BioGlyco allows the generation of specific glycoconjugates with high affinity for both academic purposes.

• Conjugation to nanoparticle carriers

Nanoparticle distribution and targeting are changed using click chemistry in vivo, thus opening new avenues for its application with in vivo drug delivery and imaging.

• Including outer membrane vesicles (OMVs), virus-like particles (VLPs), ovalbumin peptides, d-glucose, and liposomes.
• Cell labeling and tracking

The efficient labeling and precise tracking of target cells are important when monitoring biological changes such as survival, proliferation, and migration, and when evaluating the functional effects of transplanted cells in vivo.

In combination with metabolic glycoengineering, copper-free click chemistry enables easy chemical conjugation reactions to label target cells with imaging agents, such as gold nanoparticles, iron oxide nanoparticles, fluorophores, and radioisotopes, without causing severe side effects.

• Glycoconjugate characterization
• Level of chemical derivatization: Colorimetric methods, mass spectrometry (MS), and Nuclear magnetic resonance spectroscopy (NMR spectroscopy).
• Check conjugate formation: Sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE), high-performance liquid chromatography (HPLC), MS.
• Quantify different saccharide types: Colorimetric methods, high-performance anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD), and gas chromatography-mass spectrometry (GC-MS).
• Determination of glycosylated sites of protein carriers: MS.
• Identification of polysaccharide structural integrity: By immunoassays or NMR spectroscopy.

Fig.2 The diels-alder reaction-based cell surface chemical conjugation glycoengineering. (CD BioGlyco)

Applications

• The diels-alder reaction is applied to modify the chemical reporter group, such as the visualization or isolation of glycoconjugates.
• Recent application of click chemistry in drug delivery is integrating or engineering the surface of extracellular vesicles (EVs), including exosomes, which are lipid-bound vesicles (30-150 nm) that are secreted by cells.
• A click chemistry-based, two-step tumor-targeting system was used not only to deliver photosensitizers and anticancer drugs but also to deliver inhibitors.

Advantages

• The diels-alder reaction describes a class of chemical conjugation reactions that have a high reaction rate, excellent bioorthogonality, and great biocompatibility in mild aqueous conditions, orthogonal to other reactions, and which generate easily isolated products.
• The diels-alder reaction does not require a toxic catalyst, and this has consequently expanded the fields of research for which click chemistry is applicable.
• The diels-alder reaction has facilitated research on protein dynamics, animal proteome labeling, antibody engineering, and tumor pretargeting.

At CD BioGlyco, we provide high-quality Cell Surface Chemical Conjugation Glycoengineering Services for clients based on Cell Surface Glycoengineering Technology and our manufacture under a strict quality control process to ensure the client's complete satisfaction. If you are interested in our services, please feel free to contact us for more information.