Development of a multiplexed chemiluminescent immunochemical imaging technique for the simultaneous localization of different proteins in painting micro cross-sections |
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Authors: | Giorgia Sciutto Luisa Stella Dolci Angela Buragina Silvia Prati Massimo Guardigli Rocco Mazzeo Aldo Roda |
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Institution: | (1) Microchemistry and Microscopy Art Diagnostic Laboratory M2ADL, University of Bologna, Ravenna Campus, 48121 Ravenna, Italy;(2) Department of Pharmaceutical Sciences, University of Bologna, Via Belmeloro 6, 40126 Bologna, Italy; |
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Abstract: | The identification and localization of organic components in the complex stratigraphy of paintings play a crucial role in
studies of painting techniques and authentication, restoration, and conservation of artworks. Much scientific effort has been
expended for the development of analytical approaches suitable for the investigation and characterization of organic substances,
allowing high sensitivity, specificity, and spatial resolution. Proteins (e.g., ovalbumin, casein, and collagen from different
animal sources) are one of the classes of organic substances most widely used as painting materials. The analytical techniques
commonly used for their analysis (micro Fourier transform infrared spectroscopy, chromatographic techniques, and proteomic
approaches) have limits related to the lack of specificity or to the absence of information concerning the stratigraphic localization
of the detected proteins. Immunological techniques are a promising alternative approach for the characterization of proteins
in artworks. Thanks to the high specificity of antigen–antibody reactions, these techniques are widely used for the analysis
of proteins in bioanalytical and clinical chemistry and recently they have been successfully applied in the field of science
for conservation of cultural heritage. The present research aimed to develop an ultrasensitive chemiluminescent immunochemical
procedure for the simultaneous localization of ovalbumin and bovine casein (two common proteins found in binding media or
varnishes of artistic and archaeological samples) in resin-embedded painting micro cross-sections. The possibility of performing
the simultaneous identification of different proteins in painting cross-sections is of particular relevance in the field of
cultural heritage because samples are often small and available in a limited number; therefore, the maximum amount of information
must be obtained from each of them. |
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