title: A miniaturized sandwich immunoassay platform for the detection of protein-protein interactions.
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 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.
authors.affiliation: State Key Laboratory of Proteomics, Beijing Proteome Research Center, Beijing Institute of Radiation Medicine, Beijing 102206, China.
authors.forename: Qiongming
authors.initials: Q
authors.lastname: Liu
authors.forename: Qing
authors.initials: Q
authors.lastname: Chen
authors.forename: Jian
authors.initials: J
authors.lastname: Wang
authors.forename: Ying
authors.initials: Y
authors.lastname: Zhang
authors.forename: Ying
authors.initials: Y
authors.lastname: Zhou
authors.forename: Cong
authors.initials: C
authors.lastname: Lin
authors.forename: Wei
authors.initials: W
authors.lastname: He
authors.forename: Fuchu
authors.initials: F
authors.lastname: He
authors.forename: Danke
authors.initials: D
authors.lastname: Xu
mesh_terms: D057809:HEK293 Cells; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D047468:Immunoprecipitation / Q000379:methods; D040081:Protein Array Analysis / Q000379:methods; D025941:Protein Interaction Mapping / Q000379:methods*; D011506:Proteins / Q000032:analysis*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D011506:Proteins