Advanced

Integrated protein microchip assay with dual fluorescent- and MALDI read-out

Finnskog, David LU ; Ressine, Anton LU ; Laurell, Thomas LU and Marko-Varga, György LU (2004) In Journal of Proteome Research 3(5). p.988-994
Abstract
A pore chip protein array (PCPA) concept based on a dual readout configuration, fluorescence imaging, and MALDI-TOF MS has been developed. Highly packed, (>4000 Spots/cm(2)), antibody arrays were dispensed on the porous chip by using a piezo-electric microdispenser. Sandwich assay was made after blocking by addition of a secondary antibody either lgG-FITC-labeled or anti-Ang II. The antigen in the first system was a large protein (lgG), and in the other system, a FITC marked peptide Angiotensin II (Ang II) was used. Ang II antibodies showed specificity for Ang II, while the Ang I antibodies showed binding properties for Ang I, II, and Renin. Fluorescence and MALDI TOF MS read-out was made for lgG and Ang II. A major advantage of the... (More)
A pore chip protein array (PCPA) concept based on a dual readout configuration, fluorescence imaging, and MALDI-TOF MS has been developed. Highly packed, (>4000 Spots/cm(2)), antibody arrays were dispensed on the porous chip by using a piezo-electric microdispenser. Sandwich assay was made after blocking by addition of a secondary antibody either lgG-FITC-labeled or anti-Ang II. The antigen in the first system was a large protein (lgG), and in the other system, a FITC marked peptide Angiotensin II (Ang II) was used. Ang II antibodies showed specificity for Ang II, while the Ang I antibodies showed binding properties for Ang I, II, and Renin. Fluorescence and MALDI TOF MS read-out was made for lgG and Ang II. A major advantage of the dual read-out PCPA approach is that both affinity binding and mass identity are derived. Detection limits for Ang II on the chip is as low as 500 zmol (Ang II). (Less)
Please use this url to cite or link to this publication:
author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
matrix-assisted laser desorption ionization time-of-flight
in
Journal of Proteome Research
volume
3
issue
5
pages
988 - 994
publisher
The American Chemical Society
external identifiers
  • wos:000224693800009
  • scopus:7044237454
ISSN
1535-3893
DOI
10.1021/pr0499287
language
English
LU publication?
yes
id
760e48d8-3724-4cf9-8aa5-9995b4965db5 (old id 138344)
date added to LUP
2007-06-26 16:09:20
date last changed
2017-09-10 03:32:49
@article{760e48d8-3724-4cf9-8aa5-9995b4965db5,
  abstract     = {A pore chip protein array (PCPA) concept based on a dual readout configuration, fluorescence imaging, and MALDI-TOF MS has been developed. Highly packed, (>4000 Spots/cm(2)), antibody arrays were dispensed on the porous chip by using a piezo-electric microdispenser. Sandwich assay was made after blocking by addition of a secondary antibody either lgG-FITC-labeled or anti-Ang II. The antigen in the first system was a large protein (lgG), and in the other system, a FITC marked peptide Angiotensin II (Ang II) was used. Ang II antibodies showed specificity for Ang II, while the Ang I antibodies showed binding properties for Ang I, II, and Renin. Fluorescence and MALDI TOF MS read-out was made for lgG and Ang II. A major advantage of the dual read-out PCPA approach is that both affinity binding and mass identity are derived. Detection limits for Ang II on the chip is as low as 500 zmol (Ang II).},
  author       = {Finnskog, David and Ressine, Anton and Laurell, Thomas and Marko-Varga, György},
  issn         = {1535-3893},
  keyword      = {matrix-assisted laser desorption ionization time-of-flight},
  language     = {eng},
  number       = {5},
  pages        = {988--994},
  publisher    = {The American Chemical Society},
  series       = {Journal of Proteome Research},
  title        = {Integrated protein microchip assay with dual fluorescent- and MALDI read-out},
  url          = {http://dx.doi.org/10.1021/pr0499287},
  volume       = {3},
  year         = {2004},
}