Bioorthogonal chemistry

The term bioorthogonal chemistry refers to any chemical reaction that can occur inside of living systems without interfering with native biochemical processes.^[1][2][3] The term was coined by Carolyn R. Bertozzi in 2003.^[4][5] Since its introduction, the concept of the bioorthogonal reaction has enabled the study of biomolecules such as glycans, proteins,^[6] and lipids^[7] in real time in living systems without cellular toxicity. A number of chemical ligation strategies have been developed that fulfill the requirements of bioorthogonality, including the 1,3-dipolar cycloaddition between azides and cyclooctynes (also termed copper-free click chemistry),^[8] between nitrones and cyclooctynes,^[9] oxime/hydrazone formation from aldehydes and ketones,^[10] the tetrazine ligation,^[11] the isocyanide-based click reaction,^[12] and most recently, the quadricyclane ligation.^[13]

The use of bioorthogonal chemistry typically proceeds in two steps. First, a cellular substrate is modified with a bioorthogonal functional group (chemical reporter) and introduced to the cell; substrates include metabolites, enzyme inhibitors, etc. The chemical reporter must not alter the structure of the substrate dramatically to avoid affecting its bioactivity. Secondly, a probe containing the complementary functional group is introduced to react and label the substrate.

Although effective bioorthogonal reactions such as copper-free click chemistry have been developed, development of new reactions continues to generate orthogonal methods for labeling to allow multiple methods of labeling to be used in the same biosystems. Carolyn R. Bertozzi was awarded the Nobel Prize in Chemistry in 2022 for her development of click chemistry and bioorthogonal chemistry.^[14]