title: Clickable enzyme-linked immunosorbent assay.
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 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 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.
authors.affiliation: Institute for Molecules and Materials, Radboud University Nijmegen, Heyendaalseweg 135, 6525 AJ Nijmegen, The Netherlands.
authors.forename: Luiz A
authors.initials: LA
authors.lastname: Canalle
authors.forename: TuHa
authors.initials: T
authors.lastname: Vong
authors.forename: P Hans H M
authors.initials: PH
authors.lastname: Adams
authors.forename: Floris L
authors.initials: FL
authors.lastname: van Delft
authors.forename: Jos M H
authors.initials: JM
authors.lastname: Raats
authors.forename: Renato G S
authors.initials: RG
authors.lastname: Chirivi
authors.forename: Jan C M
authors.initials: JC
authors.lastname: van Hest
mesh_terms: D000480:Alkynes / Q000737:chemistry*; D000906:Antibodies / Q000276:immunology; D001172:Arthritis, Rheumatoid / Q000175:diagnosis* / Q000276:immunology; D001360:Avidin / Q000737:chemistry / Q000378:metabolism; D001386:Azides / Q000737:chemistry*; D001710:Biotin / Q000737:chemistry / Q000378:metabolism; D002384:Catalysis; D002956:Citrulline / Q000737:chemistry / Q000378:metabolism; D057930:Click Chemistry / Q000379:methods*; D003300:Copper / Q000737:chemistry; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D055767:Immobilized Proteins / Q000138:chemical synthesis / Q000276:immunology; D015394:Molecular Structure; D010455:Peptides / Q000138:chemical synthesis / Q000276:immunology
chemical_list: D000480:Alkynes; D000906:Antibodies; D001386:Azides; D055767:Immobilized Proteins; D010455:Peptides; D001360:Avidin; D002956:Citrulline; D001710:Biotin; D003300:Copper