The copper-catalyzed azide-alkyne cycloaddition reaction, or CuAAC, has been used extensively for the conjugation, immobilization, and purification of biomolecules as well as for monitoring various biological processes. Recent advances in design of copper chelating ligands such as THTPA or BTTAA improved kinetics of copper-catalyzed azide-alkyne cycloaddition reaction and greatly increased sensitivity of alkyne detection. In spite of many recent …

Figure 1). Kinetic comparison of chelating azide and non-chelating conventional azideRecent advances in the design of copper-chelating ligands, such as THPTA or BTTAA that stabilize the Cu(I) oxidation state in aqueous solution, improve the kinetics of the copper-catalyzed azide-alkyne cycloaddition (CuAAC) reaction and greatly increase the sensitivity of alkyne detection. Copper-chelating ligands have also been shown to increase the biocompatibi…

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Click-&-Go IsoTAG Kit for Profiling Intact GlycopeptidesWhile there has been much interest in profiling the intact glycoproteome, the complexity of glycoproteoforms (and more broadly, all proteoforms) remains challenging to completely define. Mass spectrometry (MS) is commonly employed for characterization of complex proteomic samples. A popular strategy for protein identification is the bottom-up shotgun proteomics approach. In this method, …

TCO – Tetrazine Ligation(Figure 1). Schematic representation of a TCO–Tz ligation reaction.At low biomolecule concentrations (e.g. < 5 μM) where SPAAC falls short due to modest kinetics (e.g. 0.3 to 2.3 M–1s–1), and where copper–catalyzed alkyne–azide cycloaddition click reaction might compromise system viability, trans–cyclooctene–tetrazine (TCO–Tz) is the reaction pair of choice. This ligation chemistry is based on an inverse–dema…

Cu – Free Click Chemistry (SPAAC)(Figure 1). Schematic representation of a SPAAC ligation reaction.The strain promoted alkyne-azide cycloaddition, also termed as the Cu-free click reaction, is a bioorthogonal reaction utilizing a pair of reagents, cyclooctynes and azides that exclusively and efficiently react with each other while remain inert to naturally occurring functional groups such as amines (Figure 1). SPAAC enables labeling a wide varie…