Clickable enzyme-linked immunosorbent assay

Article

Canalle, L.A., Vong, Tuha, Adams, P.H.H.M., van Delft, F.L., Raats, J.M.H., Chirivi, R.G.S. & Hest, van, J.C.M. (2011). Clickable enzyme-linked immunosorbent assay. Biomacromolecules, 12(10), 3692-3697. In Scopus Cited 19 times.

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Abstract

 

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 bicyclo[6.1.0]non-4-yne (BCN) chemical handle, and a diagnostic 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 ELISAs 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 immobilization methods. This technology was illustrated with an ELISA used for the diagnosis of rheumatoid arthritis (RA) but could be easily applied to a wide range of diagnostic tests.