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Issue 8, 2017
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Infrared imaging of high density protein arrays

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Abstract

We propose in this paper that protein microarrays could be analysed by infrared imaging in place of enzymatic or fluorescence labelling. This label-free method reports simultaneously a large series of data on the spotted sample (protein secondary structure, phosphorylation, glycosylation, presence of impurities, etc.). In the present work, 100 μm protein spots each containing about 100 pg protein were deposited to form high density regular arrays. Using arrays of infrared detectors, high resolution images could be obtained where each pixel of the image is in fact a full infrared spectrum. With microarrays, hundreds of experimental conditions can be tested easily and quickly, with no further labelling or chemistry of any kind. We describe how the noise present in the infrared spectra can be split into image noise and detector noise. We also detail how both types of noise can be most conveniently dealt with to generate very high quality spectra of less than 100 pg protein. Finally, the results suggest that the protein secondary structure is preserved during microarray building.

Graphical abstract: Infrared imaging of high density protein arrays

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Article information


Submitted
13 Sep 2016
Accepted
27 Nov 2016
First published
28 Nov 2016

Analyst, 2017,142, 1371-1380
Article type
Paper

Infrared imaging of high density protein arrays

J. De Meutter, J. Vandenameele, A. Matagne and E. Goormaghtigh, Analyst, 2017, 142, 1371
DOI: 10.1039/C6AN02048H

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