Institute for Molecules and Materials, Radboud University Nijmegen, Heyendaalseweg 135, 6525 AJ Nijmegen, The Netherlands.
Biomacromolecules. 2011 Oct 10;12(10):3692-7. doi: 10.1021/bm2009137. Epub 2011 Sep 1.
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.
点击化学被探索作为一种潜在的具有成本效益和选择性的固定化方法，用于生产酶联免疫吸附测定（ELISA）。制备了含有末端炔烃或双环[6.1.0]壬-4-炔（BCN）化学接头的涂层，随后通过铜催化的叠氮-炔基 1,3-偶极环加成（CuAAC）或无铜应变促进的叠氮-炔基 1,3-偶极环加成（SPAAC）分别将诊断肽固定在这些涂层上。由于固定化步骤中使用了铜催化剂，含有末端炔烃的涂层在随后的 ELISA 中显示出较高的背景水平。然而，含有 BCN 的涂层被成功地应用，并为现有的（链霉）亲和素-生物素固定化方法提供了一种具有成本效益的替代方案。该技术通过用于诊断类风湿关节炎（RA）的 ELISA 进行了说明，但可以很容易地应用于广泛的诊断测试。
