Modification of ISFETs with a monolayer of latex beads for specific detection of proteins

G.A.J. Besselink, Richardus B.M. Schasfoort, Piet Bergveld

Research output: Contribution to journalArticleAcademicpeer-review

32 Citations (Scopus)

Abstract

The so-called ion-step method is a novel potentiometric approach that can detect protein adsorbed onto the gate area of modified ion-sensitive field-effect transistors (ISFETs). In this report, a generic technology is described for immobilization of peptides and proteins to the ISFET gate in order to confer specific binding properties to the ISFET. For this, the surface of the ISFET was covered with a monolayer of Amino beads (diameter, 0.9 μm) followed by immobilization of protein ligands onto these beads. Amino beads are latex spheres that contain primary amino groups at the outer surface. Preactivation of the latex-bound amino groups with glutaraldehyde, and consecutive incubation with polylysine resulted in covalent immobilization of this polyamine, as revealed by ion stepping measurements. For ImmunoFET applications, human serum albumin (HSA) was immobilized onto the Amino bead-covered ISFETs, by passive adsorption but also by covalent coupling. Resulting devices were used for qualitative detection of α-HSA antibodies by means of the ion step method. The binding of antibody was very specific and fast (most of the binding was accomplished in 15 min) with signal yields up to 17 mV. Efforts to increase the antibody-binding capacity of the solid phase on the ISFET exploiting amino group activation (with glutaraldehyde or other homobifunctional cross linkers) before HSA coupling, did not improve signal yield. The bead technology described in this report is an easy, generic method for coating the ISFET with a solid phase that, using the ion-step method, can be applied to immunosensing.
Original languageUndefined
Pages (from-to)1109-1114
Number of pages6
JournalBiosensors and bioelectronics
Volume18
Issue number9
DOIs
Publication statusPublished - 2003

Keywords

  • ImmunoFET
  • Biosensor
  • Antibodies
  • Latex beads
  • IR-75173
  • ISFET
  • METIS-212615

Cite this