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

Författare:
Publiceringsår: 2004
Språk: Engelska
Sidor: 988-994
Publikation/Tidskrift/Serie: Journal of Proteome Research
Volym: 3
Nummer: 5
Dokumenttyp: Artikel
Förlag: American Chemical Society

Sammanfattning

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).

Disputation

Nyckelord

  • Chemistry
  • matrix-assisted laser desorption ionization time-of-flight

Övriga

Published
Yes
  • ISSN: 1535-3893

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