Clickable Enzyme-Linked Immunosorbent Assay

Click chemistry is explored as a potential cost- effective and selective immobilization method for the production of an enzyme-linked immunosorbent assay (ELISA). Coatings were formulated containing either a terminal alkyne or a bicycle[6.19]non-4-yne (BCN) chemical handle, and a diagnositc peptide was subsequently immobilized onto these coatings by the copper-catalyzed azide-alkyne 1,3-dipolar cycloaddition (CuAAC) or copper-free strain- promoted azide-alkyne 1,3-dipolar cycloaddition (SPAAC) respectively. The terminal alkyne-containing coating showed high background levels in subsequent. ELISA's due to the copper catalyst used in the immobilization step. The BCN containing coating however, was successfully employed and presents a cost-effective alternative to existing (strept) avidin-biotin immobolization methods. This technology was illustrated with an ELISA used for the diagnosis of heumatoid arthritis (RA) but could be easily applied to wide range of diagnostic tests.