RESUMO
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.
Assuntos
Alcinos/química , Artrite Reumatoide/diagnóstico , Azidas/química , Química Click/métodos , Ensaio de Imunoadsorção Enzimática , Proteínas Imobilizadas , Peptídeos , Anticorpos/imunologia , Artrite Reumatoide/imunologia , Avidina/química , Avidina/metabolismo , Biotina/química , Biotina/metabolismo , Catálise , Citrulina/química , Citrulina/metabolismo , Cobre/química , Humanos , Proteínas Imobilizadas/síntese química , Proteínas Imobilizadas/imunologia , Estrutura Molecular , Peptídeos/síntese química , Peptídeos/imunologiaRESUMO
The grafting-to of a peptide to a side chain functional polymer was investigated using "click" chemistry. Two click reactions were compared: the copper-free strain-promoted azide-alkyne 1,3-cycloaddition (SPAAC) and the traditional copper-catalyzed azide-alkyne 1,3-cycloaddition (CuAAC). For the resulting comb-shaped products, it was found that the steric bulk of the conjugation moiety used in SPAAC limits the degree of grafting for these highly dense systems, whereas CuAAC gives (near) quantitative functionalization.
Assuntos
Química Click/métodos , Gramicidina/química , Metacrilatos/química , Ácidos Polimetacrílicos/química , Alcinos/química , Azidas/química , Catálise , Cobre/química , Reação de Cicloadição , Estrutura Molecular , Triazóis/químicaRESUMO
Creating bioconjugates by combining polymers with peptides and proteins is an emerging multidisciplinary field of research that has enjoyed increased attention within the scientific community. This critical review provides an overview of the strategies employed for the construction of these materials and will highlight the underlying synthetic methods used. This review is therefore relevant for chemists, material scientists and chemical biologists facing the challenge of constructing polypeptide-polymer bioconjugates in a controlled fashion (269 references).