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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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Publication details

The article was received on 18 Dec 2012, accepted on 02 Mar 2013 and first published on 04 Mar 2013


Article type: Paper
DOI: 10.1039/C3LC41388H
Citation: Lab Chip, 2013,13, 2066-2074
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    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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