Detecting Antibodies Secreted by Trapped Cells Using Extraordinary Optical Transmission

Isolation of monoclonal (M-) antibodies (Abs) from Ab-secreting cells (ASCs) is impeded by difficulties in identifying the minority of ASCs secreting Ab(s) of interest. We present a novel label-free system that traps ASCs into biocompatible microwells and detects the binding of secreted Ab(s), to a...

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Bibliographic Details
Published in:IEEE sensors journal 2011-11, Vol.11 (11), p.2732-2739
Main Authors: Romanuik, Sean F., Grist, S. M., Gray, B. L., Hohertz, D., Kavanagh, K. L., Gulzar, N., Scott, J. K., Nirwan, R., Hui, C., Brolo, A. G., Gordon, R.
Format: Article
Language:English
Subjects:
Antibodies
Arrays
Binding
Biomedical optical imaging
Cell trap
Charge carrier processes
Exposure
extraordinary optical transmission (EOT)
Gold
immunobiosensor
Media
Nanomaterials
Nanostructure
Optical sensors
Optical surface waves
Silver
surface plasmon resonance (SPR)
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Summary:Isolation of monoclonal (M-) antibodies (Abs) from Ab-secreting cells (ASCs) is impeded by difficulties in identifying the minority of ASCs secreting Ab(s) of interest. We present a novel label-free system that traps ASCs into biocompatible microwells and detects the binding of secreted Ab(s), to a surface-immobilized antigen (Ag), as induced shifts in the extraordinary optical transmission (EOT) spectra through milled nanohole arrays. In this paper, a biotinylated target Ag (Bio-HA and Bio-2F5) is exposed to varying known MAb concentrations (17/9 and 2F5). The mean EOT shift in response to MAb concentration is similar in shape to specific MAb-Ag binding as seen in an ELISA. In another experiment, hybridoma cells secreting 17/9 MAb are trapped in wells inset into poly(dimethyl siloxane) (PDMS) (5-20 cells/trap), to which the nanohole arrays of Bio-HA "Sample" and Bio-2F5 "Ag Control" slides are aligned. Simultaneously, a Bio-HA "Media Control" slide is exposed to MAb-free media. The mean "Sample" shift (3.2 ±0.1 nm) is distinguished from the mean "Ag Control" shift (1.2 ±0.4 nm), but indistinguishable from the mean "Media Control" shift (3.6 ±0.3 nm). We have made significant progress in detecting Ab(s) secreted from trapped ASCs. With further development, our device could identify ASCs of interest rapidly, streamlining therapeutic MAb discovery.
ISSN:1530-437X
1558-1748
DOI:10.1109/JSEN.2011.2158643