Multisubstrate-compatible ELISA procedures for rapid and high-sensitivity immunoassays
Dixit, C.K. Vashist, S.K. MacCraith, B.D. O'Kennedy, R.
Dixit, C.K.
MacCraith, B.D.
O'Kennedy, R.
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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.
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Nature Protocols
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Date
2011-03
DOI
10.1038/nprot.2011.304
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Article