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- Title
A miniaturized sandwich immunoassay platform for the detection of protein-protein interactions.
- Authors
Qiongming Liu; Qing Chen; Jian Wang; Ying Zhang; Ying Zhou; Cong Lin; Wei He; Fuchu He; Danke Xu
- Abstract
Background: Analysis of protein-protein interactions (PPIs) is a valuable approach for the characterization of huge networks of protein complexes or proteins of unknown function. Co-immunoprecipitation (coIP) using affinity resins coupled to protein A/G is the most widely used method for PPI detection. However, this traditional large scale resin-based coIP is too laborious and time consuming. To overcome this problem, we developed a miniaturized sandwich immunoassay platform (MSIP) by combining antibody array technology and coIP methods. Results: Based on anti-FLAG antibody spotted aldehyde slides, MSIP enables simple, rapid and large scale detection of PPIs by fluorescent labeling anti-myc antibody. By analyzing well-known interacting and noninteracting protein pairs, MSIP was demonstrated to be highly accurate and reproducible. Compared to traditional resin-based coIP, MSIP results in higher sensitivity and enhanced throughput, with the additional benefit of digital read-outs. In addition, MSIP was shown to be a highly useful validation platform to confirm PPI candidates that have been identified from yeast two hybrid systems. Conclusions: In conclusion, MSIP is proved to be a simple, cost-saving and highly efficient technique for the comprehensive study of PPIs.
- Subjects
PROTEIN-protein interactions; IMMUNOASSAY; PROTEINS; GUMS &; resins; IMMUNOGLOBULINS
- Publication
BMC Biotechnology, 2010, Vol 10, p78
- ISSN
1472-6750
- Publication type
Article
- DOI
10.1186/1472-6750-10-78