Affordable Access

Access to the full text

Double-chip protein arrays: force-based multiplex sandwich immunoassays with increased specificity

Authors
  • Blank, Kerstin1
  • Lankenau, Andreas1
  • Mai, Thao1
  • Schiffmann, Susanne1
  • Gilbert, Ilka1
  • Hirler, Siegfried1
  • Albrecht, Christian1
  • Benoit, Martin2
  • Gaub, Hermann E.2
  • Clausen-Schaumann, Hauke1
  • 1 nanotype GmbH, Lochhamer Schlag 12, Gräfelfing, 82166, Germany , Gräfelfing
  • 2 Lehrstuhl für Angewandte Physik & Center for NanoScience, Amalienstrasse 54, München, 80799, Germany , München
Type
Published Article
Journal
Analytical and Bioanalytical Chemistry
Publisher
Springer-Verlag
Publication Date
Apr 21, 2004
Volume
379
Issue
7-8
Pages
974–981
Identifiers
DOI: 10.1007/s00216-004-2607-0
Source
Springer Nature
Keywords
License
Yellow

Abstract

Protein assays provide direct access to biologically and pharmacologically relevant information. To obtain a maximum of information from the very smallest amounts of complex biological samples, highly multiplexed protein assays are needed. However, at present, cross-reactions of binding reagents restrict the use of such assays to selected cases and severely limit the potential for up-scaling the technology. Here we describe a double-chip format, which can effectively overcome this specificity problem for sandwich immunoassays. This format consists of a capture array and a reference array with fluorescent labeled detection antibodies coupled to the reference array via DNA duplexes. This format allows for the local application of the labeled detection antibodies onto their corresponding specific spots on the capture array. Here we show that this double-chip format allows for the use of cross-reactive antibodies without generating false positive signals, and an assay for the parallel detection of seven different cytokines was set up. Even without further optimization, the dynamic range and the limit of detection for interleukin 8 were found to be comparable to those obtained with other types of multiplexed sandwich immunoassays.

Report this publication

Statistics

Seen <100 times