Heterogeneous CLICK reactions for biological conjugations
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Common CLICK reactions, often used in labeling, require ascorbic acid and organic solvents. For wider opportunity and usage in biological conjugations, CLICK reactions were performed with zinc amalgam instead of the acid. This allows for CLICK reactions to take place in virtually aqueous solutions. A simple bench-top method was created using columns of anion or cation exchange resins loaded with a copper II bathophenanthroline or copper II phendione solution, potassium phosphate buffer, and zinc amalgam. A mixture of coumarin and glycine are run through the columns and the product collected without any purification process needed. Coumarin dye was found to provide proof of a successful CLICK when the product lit up blue under UV light. High Performance Liquid Chromatography (HPLC) showed the coumarin peak in a control reaction and UV-vis indicated where the product peak should occur.