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An improved method of protein localization in artworks through SERS nanotag-complexed antibodies

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Abstract

There are several analytical techniques currently in use in conservation science to identify proteins in artworks. However, as is often the case, the determination of the exact location of a protein in a complex layer structure is challenging due to difficulty in separating layers. Localization of the protein in a cross-section has been demonstrated through attenuated total reflectance-Fourier transform infrared mapping and imaging as well as chemiluminescent and fluorescent-labeled antibodies; however, these techniques either require expensive instrumental setups or produce results that can be challenging to interpret. This paper will present research using surface-enhanced Raman scattering (SERS) nanotags complexed to secondary antibodies in conjunction with primary antibodies for the localization of ovalbumin, collagen, and casein in cross-sections from replicas and artworks containing avian egg, animal glue, or casein binders. The advantages of this technique over the others are (1) the detection method is a Raman microscope, equipment found in several museum laboratories; (2) the distinctive SERS signal from the nanotag increases the detection limit of the protein and decreases the interference from other colorants present in the cross-section layers; and finally, (3) the large (120 nm) SERS-labeled antibodies do not appear to penetrate into the cross-section, eliminating the risk of spurious signal and misidentification. Any agglomerations due to surface texture are clearly visible under normal illumination and can be avoided easily during analysis or removed with a light polish. This technique not only allows protein localization in multilayered samples while preserving the stratigraphic information but also retains the protein specificity of the antibody approach.

 

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Notes

  1. ATR-FTIR analysis performed by Adriana Rizzo, Associate Scientist, Department of Scientific Research, The Metropolitan Museum of Art, New York, USA.

  2. FTIR and GC/MS analysis performed by Adriana Rizzo, Associate Scientist, Department of Scientific Research, The Metropolitan Museum of Art, New York, USA.

  3. KPL, Kirkegaard & Perry Laboratories, Inc. online catalogue, http://www.kpl.com/catalog/categories.cfm?Catalog_ID=17&Category_ID=482

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Acknowledgments

The authors gratefully acknowledge Julia Schultz (Hochschule für Angewandte Wissenschaft und Kunst (HAWK) Fachhochschule, Hildesheim, Germany), for her generosity in allowing the reproduction of her data and the use of her methodologies in this paper and her great contribution to the immunological project at the Met during her Fellowship (2007–2009), as well as her doctoral supervisor, Dr. Karin Peterson (HAWK). The authors also thank our colleagues from the Metropolitan Museum of Art: Adriana Rizzo and Silvia Centeno, from The Department of Scientific Research, for providing samples, analytical results, and invaluable discussions; Jack Soultanian, Objects Conservator, for initiating the study of the St. John sculpture (MMA 25.120.215); and Charlotte Hale, Paintings Conservator, for launching the study of The Crucifixion (MMA 2006.409) and Scenes from the Life of St. John the Baptist (1970.134.1). Finally, we thank Dina Georgas (Barnard College, New York) for her invaluable assistance with this paper and the immunological project at the Met.

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Authors and Affiliations

  1. The Metropolitan Museum of Art, 1000 Fifth Avenue, New York, NY, 10028, USA

    Julie Arslanoglu

  2. Barnard College, 3009 Broadway, New York, NY, 10027, USA

    Stephanie Zaleski

  3. Department of Physiology and Cellular Biophysics, College of Physicians and Surgeons, Columbia University, 630 West 168 Street Room 1111, New York, NY, 10032, USA

    John Loike

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  1. Julie Arslanoglu
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Correspondence to Julie Arslanoglu.

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Published in the special issue Analytical Chemistry for Cultural Heritage with Guest Editors Rocco Mazzeo, Silvia Prati, and Aldo Roda.

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Arslanoglu, J., Zaleski, S. & Loike, J. An improved method of protein localization in artworks through SERS nanotag-complexed antibodies. Anal Bioanal Chem 399, 2997–3010 (2011). https://doi.org/10.1007/s00216-010-4378-0

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