期刊
NATURE PROTOCOLS
卷 6, 期 4, 页码 439-445出版社
NATURE PUBLISHING GROUP
DOI: 10.1038/nprot.2011.304
关键词
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资金
- Bristol-Myers Squibb (BMS), Syracuse, USA
- Industrial Development Agency, Ireland [116294]
- Science Foundation Ireland [05/CE3/B754]
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 similar to 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.
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