Abstract
Despite the large diffusion of natural organic substances in art-historical materials, their characterization presents many challenges due to the chemical complexity and instability with respect to degradation processes. Among natural products, proteins have been largely used in the past as binders but also as adhesives or additives in coating layers. Nevertheless, biological identification of proteins in art-historical objects is one of the most recent achievements obtained in heritage science thanks to the development of specifically tailored bio-analytical strategies. In the context of this active emerging discipline, immunological methods stand out for sensitivity, specificity and versatility for both protein recognition and localization in micro-samples. Furthermore, the growing use of immunological techniques for advanced diagnostics and clinical applications ensures continuous improvement in their analytical performance. Considering such, this review provides an overview of the most recent applications of enzyme linked immunosorbent assay and immunofluorescence microscopy techniques in the field of heritage materials. Specifically, the main strengths and potentials of the two techniques as well as their limits and drawbacks are presented and discussed herein.
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References
Gosling JP (1990) A decade of development in immunoassay methodology. Clin Chem 36:1408
Wild D (2013) The immunoassay handbook, 4th edn. Elsevier, Amsterdam
Gettens RJ, Stout GL (1966) Painting materials: a short encyclopaedia. Dover Publication, New York
Mills JS, White R (eds) (1994) The organic chemistry of museum objects, 2nd edn. Butterworth-Heinemann, London
Colombini MP, Modugno F (2009) Organic materials in art and archaeology. In: Colombini MP, Modugno F (eds) Organic mass spectrometry in art and archaeology, Chapter 1. Wiley, Chichester
Madariaga JM (2015) Analytical chemistry in the field of cultural heritage. Anal Methods 7:4848
Sgamellotti A, Brunetti BG, Miliani C (2014) Science and art : the painted surface. The Royal Society of Chemistry, London, UK
Domenech-Carbò MT (2008) Novel analytical methods for characterising binding media and protective coatings in artworks. Anal Chim Acta 621:109
Colombini MP, Andreotti A, Bonaduce I, Modugno F, Ribechini E (2010) Analytical strategies for characterizing organic paint media using gas chromatography/mass spectrometry. Acc Chem Res 43:715
Lluveras A, Bonaduce I, Andreotti A, Colombini MP (2010) GC/MS analytical procedure for the characterization of glycerolipids, natural waxes, terpenoid resins, proteinaceous and polysaccharide materials in the same paint microsample avoiding interferences from inorganic media. Anal Chem 8:376
Leo G, Cartechini L, Pucci P, Sgamellotti A, Marino G, Birolo L (2009) Proteomic strategies for the identification of proteinaceous binders in paintings. Anal Bioanal Chem 395:2269
Kuckova S, Hynek R, Kodicek M (2007) Identification of proteinaceous binders used in artworks by MALDI-TOF mass spectrometry. Anal Bioanal Chem 388:201
Dallongeville S, Koperska M, Garnier N, Reille-Taillefert G, Rolando C, Tokarski C (2011) Identification of animal glue species in artworks using proteomics: application to a 18th century gilt sample. Anal Chem 83:9431
Calvano CD, van der Werf ID, Palmisano F, Sabbatini L (2015) Identification of lipid- and protein-based binders in paintings by direct on-plate wet chemistry and matrix-assisted laser desorption ionization mass spectrometry. Anal Bioanal Chem 407:1015
Albertini E, Raggi L, Vagnini M, Sassolini A, Achilli A, Marconi G, Cartechini L, Veronesi F, Falcinelli M, Brunetti B, Miliani C (2011) Tracing the biological origin of animal glues used in paintings through mitochondrial DNA analysis. Anal Bioanal Chem 399:2987
Johnson M, Packard E (1971) Methods used for the identification of binding media in Italian paintings of fifteenth and sixteenth centuries. Stud Conserv 16:145
Kockaert L, Gausset P, Dubi-Rucquoy M (1989) Detection of ovalbumin in paint media by immuno-fluorescence. Stud Conserv 34:183
Raminez-Barat B, de la Vinã S (2001) Characterization of proteins in paint media by immuno-fluorescence: a note on methodological aspects. Stud Conserv 46:282
Heginbotham A, Millay V, Quick M (2006) The use of immuno-fluorescence microscopy (IFM) and enzyme-linked immunosorbent assay (ELISA) as complementary techniques for protein identification in artists’ materials. J Am Inst Conserv 45:89
Mazurek J, Heginbotham A, Schilling M, Chiari G (2008) Antibody assay to characterize binding media in paint. ICOM Comm Conserv 2:678
Cartechini L, Vagnini M, Palmieri M, Pitzurra L, Mello T, Mazurek J, Chiari G (2010) Immunodetection of proteins in ancient paint media. Acc Chem Res 43:867
Sciutto G, Dolci LS, Guardigli M, Zangheri M, Prati S, Mazzeo R, Roda A (2013) Single and multiplexed immunoassays for the chemiluminescent imaging detection of animal glues in historical paint cross-sections. Anal Bioanal Chem 405:933
Palmieri M, Vagnini M, Pitzurra L, Brunetti BG, Cartechini L (2013) Identification of animal glue and hen-egg yolk in paintings by use of enzyme-linked immunosorbent assay (ELISA). Anal Bioanal Chem 405:6365
Arslanoglu J, Zaleski S, Loike J (2011) An improved method of protein localization in artworks through SERS nanotag-complexed antibodies. Anal Bioanal Chem 399:2997
Sciutto G, Prati S, Mazzeo R, Zangheri M, Roda A, Bardini L, Valenti G, Rapino S, Marcaccio M (2014) Localization of proteins in paint cross-sections by scanning electrochemical microscopy as an alternative immunochemical detection technique. Anal Chim Acta 831:31
Lee HY, Atlasevich N, Granzotto C, Schultz J, Loike J, Arslanoglu J (2015) Development and application of an ELISA method for the analysis of protein-based binding media of artworks. Anal Methods 7:187
Magrini D, Bracci S, Sandu ICA (2013) Fluorescence of organic binders in painting cross-sections. Procedia Chem 8:194–201
Pinna D, Galleotti M, Mazzeo R (2009) Scientific examination for the investigation of paintings: a handbook for conservators-restorers. Ed. Centro Di, Firenze
Sciutto G, Litti L, Lofrumento C, Prati S, Ricci M, Gobbo M, Roda A, Castellucci E, Meneghetti M, Mazzeo R (2013) Alternative SERRS probes for the immunochemical localization of ovalbumin in paintings: an advanced mapping detection approach. Analyst 138:4532–4541
Perets EA, Indrasekara ASDS, Kurmis A, Atlasevich N, Fabris L, Arslanoglu J (2015) Carboxy-terminated immuno-SERS tags overcome non-specific aggregation for the robust detection and localization of organic media in artworks. Analyst 140:5971–5980
Avci R, Schweitzer MH, Boyd RD, Wittmeyer JL, Terán Arce F, Calvo JO (2005) Preservation of bone collagen from the late cretaceous period studied by immunological techniques and atomic force microscopy. Langmuir 21:3584–3590
Lindgren J, Uvdal P, Engdahl A, Lee AH, Alwmark C, Bergquist C-E, Nilsson E, Ekström P, Rasmussen M, Douglas DA, Polcyn MJ, Jacobs LL (2011) Microspectroscopic evidence of cretaceous bone proteins. PLoS ONE 6(4):e19445
Vagnini M, Pitzurra L, Cartechini L, Miliani C, Brunetti BG, Sgamellotti A (2008) Identification of proteins in painting cross-sections by immunofluorescence microscopy. Anal Bioanal Chem 392:57–64
Klausmeyer PA, Albertson RP, Woodland RT, Schmidt MR, Blewett M (2009) FTIR and ELISA for the analysis of a Kees Van Dongen painting. e-PS 6:151
Scott DA, Warmlander S, Mazurek J, Quirke S (2009) Examination of some pigments, grounds and media from Egyptian cartonnage fragments in the Petrie Museum, University College London. J Archaeol Sci 36:923–932
Palmieri M, Vagnini M, Pitzurra L, Rocchi P, Brunetti BG, Sgamellotti A, Cartechini L (2011) Development of an analytical protocol for a fast, sensitive and specific protein recognition in paintings by enzyme-linked immunosorbent assay (ELISA). Anal Bioanal Chem 399:3011–3023
Dolci LS, Sciutto G, Guardigli A, Rizzoli M, Prati S, Mazzeo R, Roda A (2008) Ultrasensitive chemiluminescent immunochemical identification and localization of protein components in painting cross-sections by microscope low-light imaging. Anal Bioanal Chem 392:29–35
Sciutto G, Dolci LS, Buragina A, Prati S, Guardigli A, Mazzeo R, Roda A (2011) Development of a multiplexed chemiluminescent immunochemical imaging technique for the simultaneous localization of different proteins in painting micro cross-sections. Anal Bioanal Chem 399:2889–2897
Zangheri M, Sciutto G, Mirasoli M, Prati S, Mazzeo R, Roda A, Guardigli M (2016) A portable device for on site detection of chicken ovalbumin in artworks by chemiluminescent immunochemical contact imaging. Microchem J 124:247–255
Mazurek J, Svoboda M, Maish J, Kawahara K, Fukakusa S, Nakazawa T, Taniguchi Y (2014) Characterization of binding media in Egyptian Romano portraits using enzyme-linked immunosorbent assay and mass spectrometry. e-PS 11:76
Hu W, Zhang K, Zhang H, Zhang B, Rong B (2015) Analysis of polychromy binder on Qin Shihuang’s terracotta warriors by immunofluorescence microscopy. J Cult Herit 16:244
Gambino M, Cappitelli F, Cattò C, Carpen A, Principi P, Ghezzi L, Bonaduce I, Galano E, Pucci P, Birolo L, Villa F, Forlani F (2013) A simple and reliable methodology to detect egg white in art samples. J Biosci 38:397
Potenza M, Sabatino G, Giambi F, Rosi L, Papini AM, Dei L (2013) Analysis of egg-based model wall paintings by use of an innovative combined dot-ELISA and UPLC-based approach. Anal Bioanal Chem 405:691
Bottari F, Oliveri P, Ugo P (2014) Electrochemical immunosensor based on ensemble of nanoelectrodes for immunoglobulin IgY detection: application to identify hen’s egg yolk in tempera paintings. Biosens Bioelectron 52:403
Zheng Q, Wu X, Zheng H, Zhou Y (2015) Development of an enzyme-linked-immunosorbent-assay technique for accurate identification of poorly preserved silks unearthed in ancient tombs. Anal Bioanal Chem 407:3861–3867
Sela-Culang I, Kunik V, Ofran Y (2013) The structural basis of antibody-antigen recognition. Front Immunol 4:302. doi:10.3389/fimmu.2013.00302
Akiba H, Tsumoto K (2015) Thermodynamics of antibody-antigen interaction revealed by mutation analysis of antibody variable regions. J Biochem 158:1–13
Moreira IS, Fernandes PA, Ramos MJ (2007) Hot spots—a review of the protein–protein interface determinant amino-acid residues. Proteins 68:803–812
Borrebaeck CAK (2000) Antibodies in diagnostics—from immunoassays to protein chips. Immunol Today 21:379
Hage DS (1995) Immunoassays. Anal Chem 67:455R
Blake C, Gould BJ (1984) Use of enzymes in immunoassay techniques. A review. Analyst 109:53
Porstmann T, Kiessig ST (1992) Enzyme immunoassay techniques. An overview. J Immunol Methods 150:5
Petty HR (2007) Fluorescence microscopy: established and emerging methods, experimental strategies, and applications in immunology. Microsc Res Tech 70:687
Lequin RM (2005) Enzyme immunoassay (EIA)/enzyme-linked immunosorbent assay (ELISA). Clin Chem 51:2415–2418
Crowther JR (2009) The ELISA guidebook, 2nd edn. Humana Press, Totowa, NJ
Giepmans BNG, Adams SR, Ellisman MH, Tsien RY (2006) The fluorescent toolbox for assessing protein location and function. Science 312:217
Resch-Genger U, Grabolle M, Cavaliere-Jaricot S, Nitschke R, Nann T (2008) Quantum dots versus organic dyes as fluorescent labels. Nat Methods 5:763–775
Rino J, Braga J, Henriques R, Carmo-Fonseca M (2009) Frontiers in fluorescence microscopy Int. J Dev Biol 53:1569
Schermelleh L, Heintzmann R, Leonhardt H (2010) A guide to super-resolution fluorescence microscopy. J Cell Biol 190:165
Colombini MP, Modugno F (2004) Characterisation of proteinaceous binders in artistic paintings by chromatographic techniques. J Sep Sci 27:147
Lai MC, Topp EM (1999) Solid-state chemical stability of proteins and peptides. J Pharm Sci 88:489–500
Karpowicz A (1981) Ageing and deterioration of proteinaceous media. Stud Conserv 26:153–160
Bonaduce I, Cito M, Colombini MP (2009) The development of a gas chromatographic–mass spectrometric analytical procedure for the determination of lipids, proteins and resins in the same paint micro-sample avoiding interferences from inorganic media. J Chromatogr A 1216:5931
Leo G, Bonaduce I, Andreotti A, Marino G, Pucci P, Colombini MP, Birolo L (2011) Deamidation at asparagine and glutamine as a major modification upon deterioration/aging of proteinaceous binders in mural paintings. Anal Chem 83:2056
Duce C, Bramanti E, Ghezzi L, Bernazzani L, Bonaduce I, Colombini MP, Spepi A, Biagi S, Tine MR (2013) Interactions between inorganic pigments and proteinaceous binders in reference paint reconstructions. Dalton Trans 42:5975
Ghezzi L, Duce C, Bernazzani L, Bramanti E, Colombini MP, Tiné MR, Bonaduce I (2015) Interactions between inorganic pigments and rabbit skin glue in reference paint reconstructions. J Therm Anal Calorim 122:315–322
Arslanoglu J, Schultz J, Loike J, Peterson K (2010) Immunology and art: using antibody-based techniques to identify proteins and gums in artworks. J Biosci 35:3
Ren F, Atlasevich N, Baade B, Loike J, Arslanoglu J (2015) Influence of pigments and protein aging on protein identification in historically representative casein-based paints using enzyme-linked immunosorbent assay. Anal Bioanal Chem. doi:10.1007/s00216-015-9089-0
Zevgiti S, Sackarellos C, Sackarellos-Daitsiotis M, Ioakimoglou E, Panou-Pomonis E (2007) Collagen models as a probe in the decay of works of art: synthesis, conformation and immunological studies. J Pept Sci 13:121–127
Brunello F (1982) Il libro dell’arte, Neri, Pozza edn. Vicenza, Italy
Ishikawa E, Hashida S, Kohno T (1991) Development of ultrasensitive enzyme immunoassay reviewed with emphasis on factors which limit the sensitivity. Mol Cell Probes 5:81–95
Towbin H, Gordon J (1984) Immunoblotting and dot immunobinding—current status and outlook. J Immunol Method 72:313–340
Lichtman JW, Conchello JA (2005) Fluorescence microscopy. Nat Methods 2:910–919
Matteini P, Camaiti M, Agati G, Baldo MA, Mutoc S, Matteini M (2009) Discrimination of painting binders subjected to photo-ageing by using microspectrofluorometry coupled with deconvolution analysis. J Cult Herit 10:198–205
Nevin A, Anglos D, Cather S, Burnstock A (2008) The influence of visible light and inorganic pigments on fluorescence excitation emission spectra of egg-, casein- and collagen-based painting media. Appl Phys A 92:69–76
Sandu ICA, Roque ACA, Matteini P, Schaefer S, Aagati G, Correia CR, Viana JFFP (2012) Fluorescence recognition of proteinaceous binders in works of art by a novel integrated system of investigation. Microsc Microanal 75:316–324
Sandu ICA, Schaefer S, Magrini D, Bracci S, Roque ACA (2012) Cross-section and staining-based techniques for investigation of organic materials in painted and polychrome works of art—a review. Microsc Microanal 18:860–865
Földes-Papp Z, Demel U, Tilz GP (2003) Laser scanning confocal fluorescence microscopy: an overview. Int Immunopharmacol 3:1715–1729
Panchuk-Voloshina N, Haugland RP, Bishop-Stewart J, Bhalgat MK, Millard PJ, Mao F, Leung W-Y, Haugland RP (1999) Alexa Dyes, a series of new fluorescent dyes that yield exceptionally bright, photostable conjugates. J Histochem Cytochem 47:1179–1188
Sarkar P, Sridharan S, Luchowski R, Desai S, Dworecki B, Nlend M, Gryczynski Z, Gryczynski I (2010) Photophysical properties of a new DyLight 594 dye. J Photochem Photobiol, B 98:35–39
Schermelleh L, Heintzmann R, Leonhardt H (2010) A guide to super-resolution fluorescence microscopy. J Cell Biol 190:165–175
Pavelka J, Kovaciková L, Smejda L (2011) The determination of domesticated animal species from a Neolithic sample using the ELISA test. C R Palevol 10:61–70
Yao J, Yang M, Duan Y (2014) Chemistry, biology, and medicine of fluorescent nanomaterials and related systems: new insights into biosensing, bioimaging, genomics, diagnostics, and therapy. Chem Rev 114:6130–6178
Day JJ, Marquez BV, Beck HE, Aweda TA, Gawande PD, Meares CF (2010) Chemically modified antibodies as diagnostic imaging agents. Curr Opin Chem Biol 14:803–809
Beltran V, Salvadó N, Butí S, Cinque G, Wehbe K, Pradell T (2015) Optimal sample preparation for the analysis of micrometric heterogeneous samples. Anal Chem 87:6500–6504
Acknowledgments
The authors acknowledge for funding support the CHARISMA project (GA228330)—funded by the European Union FP7-Research Infrastructure programme—and the project “Sviluppo delle attività di ricerca, valutazione e tutela conservative” of the Regione Umbria—“Progetto 1 del Primo atto integrativo all’APQ: Tutela e prevenzione dei beni culturali”.
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Cartechini, L., Palmieri, M., Vagnini, M. et al. Immunochemical Methods Applied to Art-Historical Materials: Identification and Localization of Proteins by ELISA and IFM. Top Curr Chem (Z) 374, 5 (2016). https://doi.org/10.1007/s41061-015-0006-y
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DOI: https://doi.org/10.1007/s41061-015-0006-y