A comparative analysis of polyurethane hydrogel for immobilization of IgG on chips

Katarzyna Derwinska, Levi A. Gheber, Claudia Preininger

Research output: Contribution to journalArticlepeer-review

25 Scopus citations

Abstract

Hydrogels are considered an optimum material for protein chip surfaces, since they provide a quasi-liquid environment which allows protein activity to be maintained and shows good spot morphology as well as excellent immobilization capacity. In the following, we present a polyurethane (PU) chip that electrostatically binds IgG. The PU surface is optimized with regard to layer thickness (∼200 nm), hydrogel (2%) and immobilized antibody concentration (0.5 mg mL-1; 0.3 ng spot-1), pH and ionic strength of the print buffer as well as to blocking solution. Evaluation is done in a direct IgG immunoassay using the Nexterion slide H as a reference. It is shown that higher IgG loading is achieved on the PU chip than on slide H, no matter whether 1× PBS (pH 7.2), Sörensen (pH 5.8) or Nexterion buffer was used as a spotting solution. Moreover, the crossreactivity with goat IgG, human IgG and monoclonal anti-CRP spotted in Nexterion buffer was as low as ≤0.74% (slide H: ≤3.34%).

Original languageEnglish
Pages (from-to)132-138
Number of pages7
JournalAnalytica Chimica Acta
Volume592
Issue number2
DOIs
StatePublished - 5 Jun 2007

Keywords

  • Antibody immobilization
  • Polyurethane
  • Print buffer
  • Protein microarray

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Environmental Chemistry
  • Spectroscopy

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