Abstract
In solid-support immunoassays, the transport of target analyte in sample solution to capture molecules on the sensor surface controls the detected binding signal. Depletion of the target analyte in the sample solution adjacent to the sensor surface leads to deviations from ideal association, and causes inhomogeneity of surface binding as analyte concentration varies spatially across the sensor surface. In the field of label-free optical biosensing, studies of mass-transport-limited reaction kinetics have focused on the average response on the sensor surface, but have not addressed binding inhomogeneities caused by mass-transport limitations. In this paper, we employ Molecular Interferometric Imaging (MI2) to study mass-transport-induced inhomogeneity of analyte binding within a single protein spot. Rabbit IgG binding to immobilized protein A/G was imaged at various concentrations and under different flow rates. In the mass-transport-limited regime, enhanced binding at the edges of the protein spots was caused by depletion of analyte towards the center of the protein spots. The magnitude of the inhomogeneous response was a function of analyte reaction rate and sample flow rate.
| Original language | English (US) |
|---|---|
| Title of host publication | Progress in Biomedical Optics and Imaging - Proceedings of SPIE |
| Volume | 7188 |
| DOIs | |
| State | Published - 2009 |
| Externally published | Yes |
| Event | Nanoscale Imaging, Sensing, and Actuation for Biomedical Applications VI - San Jose, CA, United States Duration: Jan 27 2009 → Jan 28 2009 |
Other
| Other | Nanoscale Imaging, Sensing, and Actuation for Biomedical Applications VI |
|---|---|
| Country | United States |
| City | San Jose, CA |
| Period | 1/27/09 → 1/28/09 |
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ASJC Scopus subject areas
- Atomic and Molecular Physics, and Optics
- Electronic, Optical and Magnetic Materials
- Biomaterials
- Radiology Nuclear Medicine and imaging
Cite this
Probing mass-transport and binding inhomogeneity in macromolecular interactions by molecular interferometric imaging. / Zhao, Ming; Wang, Xuefeng; Nolte, David.
Progress in Biomedical Optics and Imaging - Proceedings of SPIE. Vol. 7188 2009. 71880O.Research output: Chapter in Book/Report/Conference proceeding › Conference contribution
}
TY - GEN
T1 - Probing mass-transport and binding inhomogeneity in macromolecular interactions by molecular interferometric imaging
AU - Zhao, Ming
AU - Wang, Xuefeng
AU - Nolte, David
PY - 2009
Y1 - 2009
N2 - In solid-support immunoassays, the transport of target analyte in sample solution to capture molecules on the sensor surface controls the detected binding signal. Depletion of the target analyte in the sample solution adjacent to the sensor surface leads to deviations from ideal association, and causes inhomogeneity of surface binding as analyte concentration varies spatially across the sensor surface. In the field of label-free optical biosensing, studies of mass-transport-limited reaction kinetics have focused on the average response on the sensor surface, but have not addressed binding inhomogeneities caused by mass-transport limitations. In this paper, we employ Molecular Interferometric Imaging (MI2) to study mass-transport-induced inhomogeneity of analyte binding within a single protein spot. Rabbit IgG binding to immobilized protein A/G was imaged at various concentrations and under different flow rates. In the mass-transport-limited regime, enhanced binding at the edges of the protein spots was caused by depletion of analyte towards the center of the protein spots. The magnitude of the inhomogeneous response was a function of analyte reaction rate and sample flow rate.
AB - In solid-support immunoassays, the transport of target analyte in sample solution to capture molecules on the sensor surface controls the detected binding signal. Depletion of the target analyte in the sample solution adjacent to the sensor surface leads to deviations from ideal association, and causes inhomogeneity of surface binding as analyte concentration varies spatially across the sensor surface. In the field of label-free optical biosensing, studies of mass-transport-limited reaction kinetics have focused on the average response on the sensor surface, but have not addressed binding inhomogeneities caused by mass-transport limitations. In this paper, we employ Molecular Interferometric Imaging (MI2) to study mass-transport-induced inhomogeneity of analyte binding within a single protein spot. Rabbit IgG binding to immobilized protein A/G was imaged at various concentrations and under different flow rates. In the mass-transport-limited regime, enhanced binding at the edges of the protein spots was caused by depletion of analyte towards the center of the protein spots. The magnitude of the inhomogeneous response was a function of analyte reaction rate and sample flow rate.
UR - http://www.scopus.com/inward/record.url?scp=66649134674&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=66649134674&partnerID=8YFLogxK
U2 - 10.1117/12.809500
DO - 10.1117/12.809500
M3 - Conference contribution
AN - SCOPUS:66649134674
SN - 9780819474346
VL - 7188
BT - Progress in Biomedical Optics and Imaging - Proceedings of SPIE
ER -