Mass-transport limitations in spot-based microarrays

Ming Zhao, Xuefeng Wang, David Nolte

Research output: Contribution to journalArticle

9 Scopus citations

Abstract

Mass transport of analyte to surface-immobilized affinity reagents is the fundamental bottleneck for sensitive detection in solidsupport microarrays and biosensors. Analyte depletion in the volume adjacent to the sensor causes deviation from ideal association, significantly slows down reaction kinetics, and causes inhomogeneous binding across the sensor surface. In this paper we use high-resolution molecular interferometric imaging (MI2), a label-free optical interferometry technique for direct detection of molecular films, to study the inhomogeneous distribution of intra-spot binding across 100 micron-diameter protein spots. By measuring intra-spot binding inhomogeneity, reaction kinetics can be determined accurately when combined with a numerical three-dimensional finite element model. To ensure homogeneous binding across a spot, a critical flow rate is identified in terms of the association rate ka and the spot diameter. The binding inhomogeneity across a spot can be used to distinguish high-affinity low-concentration specific reactions from lowaffinity high-concentration non-specific binding of background proteins.

Original languageEnglish (US)
Pages (from-to)983-997
Number of pages15
JournalBiomedical Optics Express
Volume1
Issue number3
DOIs
StatePublished - Oct 1 2010

ASJC Scopus subject areas

  • Biotechnology
  • Atomic and Molecular Physics, and Optics

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