Mass-transport limitations in spot-based microarrays

Ming Zhao, Xuefeng Wang, David Nolte

Research output: Contribution to journalArticle

9 Citations (Scopus)

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

Fingerprint

Interferometry
Molecular Imaging
Biosensing Techniques
Carrier Proteins
affinity
inhomogeneity
reaction kinetics
critical flow
proteins
Proteins
causes
sensors
bioinstrumentation
reagents
low concentrations
interferometry
depletion
flow velocity
deviation
high resolution

ASJC Scopus subject areas

  • Atomic and Molecular Physics, and Optics
  • Biotechnology

Cite this

Mass-transport limitations in spot-based microarrays. / Zhao, Ming; Wang, Xuefeng; Nolte, David.

In: Biomedical Optics Express, Vol. 1, No. 3, 01.10.2010, p. 983-997.

Research output: Contribution to journalArticle

Zhao, Ming ; Wang, Xuefeng ; Nolte, David. / Mass-transport limitations in spot-based microarrays. In: Biomedical Optics Express. 2010 ; Vol. 1, No. 3. pp. 983-997.
@article{6cffc6348fff4dd9a56fcadcfcca1676,
title = "Mass-transport limitations in spot-based microarrays",
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.",
author = "Ming Zhao and Xuefeng Wang and David Nolte",
year = "2010",
month = "10",
day = "1",
doi = "10.1364/BOE.1.000983",
language = "English (US)",
volume = "1",
pages = "983--997",
journal = "Biomedical Optics Express",
issn = "2156-7085",
publisher = "The Optical Society",
number = "3",

}

TY - JOUR

T1 - Mass-transport limitations in spot-based microarrays

AU - Zhao, Ming

AU - Wang, Xuefeng

AU - Nolte, David

PY - 2010/10/1

Y1 - 2010/10/1

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=84862891287&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84862891287&partnerID=8YFLogxK

U2 - 10.1364/BOE.1.000983

DO - 10.1364/BOE.1.000983

M3 - Article

AN - SCOPUS:84862891287

VL - 1

SP - 983

EP - 997

JO - Biomedical Optics Express

JF - Biomedical Optics Express

SN - 2156-7085

IS - 3

ER -