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Issue 11, 2013
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Scaling advantages and constraints in miniaturized capture assays for single cell protein analysis

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Abstract

Measuring protein expression in single cells is the basis of single cell proteomics. The sensitivity and dynamic range of a single cell immunoassay should ideally be such that proteins that are expressed between 1–106 copies per cell can be detected and counted. We have investigated the effect of miniaturizing antibody microarrays by reducing capture spot sizes from 100 μm to 15 μm using dip-pen nanolithography. We demonstrate that protocols developed for printing and passivating antibody capture spots using conventional pin-based contact printing can be directly transferred to dip-pen lithography whilst retaining the capture activity per unit area. Using a simple kinetic model, we highlight how the limit of detection and dynamic range of a sandwich immunoassay, respectively, increase and decrease when spot size is reduced. However, we show that reducing spot size is more effective than increasing assay chamber volume when seeking to multiplex such a microfluidic immunoassay. Although we make particular reference to single cell microfluidic immunoassays, the topics discussed here are applicable to capture assays in general.

Graphical abstract: Scaling advantages and constraints in miniaturized capture assays for single cell protein analysis

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Submitted
18 Dec 2012
Accepted
02 Mar 2013
First published
04 Mar 2013

Lab Chip, 2013,13, 2066-2074
Article type
Paper

Scaling advantages and constraints in miniaturized capture assays for single cell protein analysis

A. Salehi-Reyhani, S. Sharma, E. Burgin, M. Barclay, A. Cass, M. A. A. Neil, O. Ces, K. R. Willison and D. R. Klug, Lab Chip, 2013, 13, 2066
DOI: 10.1039/C3LC41388H

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