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A miniaturized sandwich immunoassay platform for the detection of protein-protein interactions
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 detecti...
Autores principales: | , , , , , , , , |
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Formato: | Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2010
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2978116/ https://www.ncbi.nlm.nih.gov/pubmed/20979660 http://dx.doi.org/10.1186/1472-6750-10-78 |
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author | Liu, Qiongming Chen, Qing Wang, Jian Zhang, Ying Zhou, Ying Lin, Cong He, Wei He, Fuchu Xu, Danke |
author_facet | Liu, Qiongming Chen, Qing Wang, Jian Zhang, Ying Zhou, Ying Lin, Cong He, Wei He, Fuchu Xu, Danke |
author_sort | Liu, Qiongming |
collection | PubMed |
description | 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 non-interacting 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. |
format | Text |
id | pubmed-2978116 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2010 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-29781162010-11-17 A miniaturized sandwich immunoassay platform for the detection of protein-protein interactions Liu, Qiongming Chen, Qing Wang, Jian Zhang, Ying Zhou, Ying Lin, Cong He, Wei He, Fuchu Xu, Danke BMC Biotechnol Methodology Article 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 non-interacting 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. BioMed Central 2010-10-28 /pmc/articles/PMC2978116/ /pubmed/20979660 http://dx.doi.org/10.1186/1472-6750-10-78 Text en Copyright ©2010 Liu et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Methodology Article Liu, Qiongming Chen, Qing Wang, Jian Zhang, Ying Zhou, Ying Lin, Cong He, Wei He, Fuchu Xu, Danke A miniaturized sandwich immunoassay platform for the detection of protein-protein interactions |
title | A miniaturized sandwich immunoassay platform for the detection of protein-protein interactions |
title_full | A miniaturized sandwich immunoassay platform for the detection of protein-protein interactions |
title_fullStr | A miniaturized sandwich immunoassay platform for the detection of protein-protein interactions |
title_full_unstemmed | A miniaturized sandwich immunoassay platform for the detection of protein-protein interactions |
title_short | A miniaturized sandwich immunoassay platform for the detection of protein-protein interactions |
title_sort | miniaturized sandwich immunoassay platform for the detection of protein-protein interactions |
topic | Methodology Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2978116/ https://www.ncbi.nlm.nih.gov/pubmed/20979660 http://dx.doi.org/10.1186/1472-6750-10-78 |
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