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|Title:||Multisubstrate-compatible ELISA procedures for rapid and high-sensitivity immunoassays|
|Citation:||Dixit, C.K., Vashist, S.K., MacCraith, B.D., O'Kennedy, R. (2011-03). Multisubstrate-compatible ELISA procedures for rapid and high-sensitivity immunoassays. Nature Protocols 6 (4) : 439-445. ScholarBank@NUS Repository. https://doi.org/10.1038/nprot.2011.304|
|Abstract:||This protocol describes an improved and optimized approach to develop rapid and high-sensitivity ELISAs by covalently immobilizing antibody on chemically modified polymeric surfaces. The method involves initial surface activation with KOH and an O 2 plasma, and then amine functionalization with 3-aminopropyltriethoxysilane. The second step requires covalent antibody immobilization on the aminated surface, followed by ELISA. The ELISA procedure developed is 16-fold more sensitive than established methods. This protocol could be used generally as a quantitative analytical approach to perform high-sensitivity and rapid assays in clinical situations, and would provide a faster approach to screen phage-displayed libraries in antibody development facilities. The antibody immobilization procedure is of ∼3 h duration and facilitates rapid ELISAs. This method can be used to perform assays on a wide range of commercially relevant solid support matrices (including those that are chemically inert) with various biosensor formats. © 2011 Nature America, Inc. All rights reserved.|
|Source Title:||Nature Protocols|
|Appears in Collections:||Staff Publications|
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