A miniaturized sandwich immunoassay platform for the detection of protein-protein interactions
<p>Abstract</p> <p>Background</p> <p>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 pro...
Main Authors: | , , , , , , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
BMC
2010-10-01
|
Series: | BMC Biotechnology |
Online Access: | http://www.biomedcentral.com/1472-6750/10/78 |
id |
doaj-8a4c42e434264182bc06f67b174519c1 |
---|---|
record_format |
Article |
spelling |
doaj-8a4c42e434264182bc06f67b174519c12020-11-25T01:38:55ZengBMCBMC Biotechnology1472-67502010-10-011017810.1186/1472-6750-10-78A miniaturized sandwich immunoassay platform for the detection of protein-protein interactionsHe WeiLin CongZhou YingZhang YingWang JianChen QingLiu QiongmingHe FuchuXu Danke<p>Abstract</p> <p>Background</p> <p>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.</p> <p>Results</p> <p>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.</p> <p>Conclusions</p> <p>In conclusion, MSIP is proved to be a simple, cost-saving and highly efficient technique for the comprehensive study of PPIs.</p> http://www.biomedcentral.com/1472-6750/10/78 |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
He Wei Lin Cong Zhou Ying Zhang Ying Wang Jian Chen Qing Liu Qiongming He Fuchu Xu Danke |
spellingShingle |
He Wei Lin Cong Zhou Ying Zhang Ying Wang Jian Chen Qing Liu Qiongming He Fuchu Xu Danke A miniaturized sandwich immunoassay platform for the detection of protein-protein interactions BMC Biotechnology |
author_facet |
He Wei Lin Cong Zhou Ying Zhang Ying Wang Jian Chen Qing Liu Qiongming He Fuchu Xu Danke |
author_sort |
He Wei |
title |
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_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_sort |
miniaturized sandwich immunoassay platform for the detection of protein-protein interactions |
publisher |
BMC |
series |
BMC Biotechnology |
issn |
1472-6750 |
publishDate |
2010-10-01 |
description |
<p>Abstract</p> <p>Background</p> <p>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.</p> <p>Results</p> <p>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.</p> <p>Conclusions</p> <p>In conclusion, MSIP is proved to be a simple, cost-saving and highly efficient technique for the comprehensive study of PPIs.</p> |
url |
http://www.biomedcentral.com/1472-6750/10/78 |
work_keys_str_mv |
AT hewei aminiaturizedsandwichimmunoassayplatformforthedetectionofproteinproteininteractions AT lincong aminiaturizedsandwichimmunoassayplatformforthedetectionofproteinproteininteractions AT zhouying aminiaturizedsandwichimmunoassayplatformforthedetectionofproteinproteininteractions AT zhangying aminiaturizedsandwichimmunoassayplatformforthedetectionofproteinproteininteractions AT wangjian aminiaturizedsandwichimmunoassayplatformforthedetectionofproteinproteininteractions AT chenqing aminiaturizedsandwichimmunoassayplatformforthedetectionofproteinproteininteractions AT liuqiongming aminiaturizedsandwichimmunoassayplatformforthedetectionofproteinproteininteractions AT hefuchu aminiaturizedsandwichimmunoassayplatformforthedetectionofproteinproteininteractions AT xudanke aminiaturizedsandwichimmunoassayplatformforthedetectionofproteinproteininteractions AT hewei miniaturizedsandwichimmunoassayplatformforthedetectionofproteinproteininteractions AT lincong miniaturizedsandwichimmunoassayplatformforthedetectionofproteinproteininteractions AT zhouying miniaturizedsandwichimmunoassayplatformforthedetectionofproteinproteininteractions AT zhangying miniaturizedsandwichimmunoassayplatformforthedetectionofproteinproteininteractions AT wangjian miniaturizedsandwichimmunoassayplatformforthedetectionofproteinproteininteractions AT chenqing miniaturizedsandwichimmunoassayplatformforthedetectionofproteinproteininteractions AT liuqiongming miniaturizedsandwichimmunoassayplatformforthedetectionofproteinproteininteractions AT hefuchu miniaturizedsandwichimmunoassayplatformforthedetectionofproteinproteininteractions AT xudanke miniaturizedsandwichimmunoassayplatformforthedetectionofproteinproteininteractions |
_version_ |
1725051369312747520 |