A New Heterogeneous Method of CLICK Reactions for Biological Conjugations
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The simple and fast CLICK reaction, in which an azide is linked to an alkyne, has been used extensively to produce substances promptly and reliably. These reactions commonly require ascorbic acid and organic solvents, limiting opportune application. Substituting zinc amalgam for the acid provides a heterogeneous system in which the reductant can easily be separated from the desired product. This system of reduction also allows for the CLICK to occur in virtually aqueous solutions. Evidence of this successful practicality was obtained by CLICKing an azide labeled virus to an alkyne labeled fluorescein dye, using a copper II bathophenanthroline catalyst, Tris pH 8 buffer and zinc amalgam. The zinc amalgam was added to a solution of copper II bathophenanthroline to reduce the copper from copper II to the copper I needed for the CLICK. After the solution has been altered from a turquoise color to a deep shade of green, indicating the reduction, the virus in buffer and fluorescein in dimethyl sulfoxide (DMSO) were added. The product was verified by comparison to a successful ascorbate CLICK of identical substrates run on a protein gel.