Final Report Summary - NANOART (Nano Art Research Tool)
The characterization of historic paints’ binders still relies on conventional molecular biology methodologies that were developed decades ago and which have been being substituted by more sensitive, specific, inexpensive and faster methodologies, taking advantage of the benefits of the emerging nanotechnology world. Therefore, the NANOART project was built to innovate by transferring and upgrading these methodologies with top-notch technology that is being developed for the area of clinical diagnostic. In this sense, the project was original, since it took the latest developments carried out for the wealthy area of medicine in benefit of the Conservation and preservation of Cultural Heritage area, further lowering the costs of research and maximizing synergies, creating the NANOART kit, where the final user will be able to conduct an affordable analysis (in most cases it costs only a few euros) into lower quantities of sample from the artwork, by just reading some instructions and within a 4 hour period will have the results.
Through the research of Medieval and Renaissance treatises, referring the manufacturing process of paints, such as: On Divers Arts (12th century), Il libro del art (14th century), The Strasburg Manuscript (late Middle Ages), The book on how to make colours (a late medieval Portuguese technical text on illumination), 5) Vasari on Technique (1550) and The B.M. Sloane 2052 (17th century), three classes of binders were identified: proteinaceous, glycosides and oil binders. Dr Ana Claro decided to start with the proteinaceous ones, meaning with animal glue, mainly composed by collagen and egg white.
After an exhaustive research two antibodies for collagen type I, and two for ovalbumin (mono and polyclonal), were selected, to further on build up the NANOKit.
Having the biomarkers selected, standard samples mimicking real samples were made (Figures 1 and 2) and the development of the protocol that would establish the way on how the samples should be treated previously to the detection was created.
Synthesis and functionalization of Gold Nanoparticles (GNPs) were fundamental steps since it provided the nanoparticles necessary for the nanobiosensor. Once synthesized, the GNPs were functionalized with the different targeting molecules, by following the functionalization methodologies developed by the NG2. These targeting molecules were covalently attached to the nanoparticles’ surface and in an oriented way, so they can successfully bind to their target molecules, giving rise to GNPs conjugates, also known as gold nanoprobes.
The methodology of the nanobiosensor used in this project has been developed by NG2: the photothermal nanobiosensor (Figure 3). This sensor is based on GNPs conjugates that generate heat upon being irradiated with a laser. This method is more sensitive than the colorimetric one (first one selected for the NANOART project) although requires a more complex mechanism to reveal the final result. This issue was solved with the development of a hard and software for the laser set up (Figures 4 and 5).
Analyzes performed with the model samples showed in most of the cases the expected results. Some exceptions occurred when an interaction (expected and reported in literature) between the pigments and the binder could happen, namely with minium, cinnabar and white lead. Knowing that one can recommend microsampling stable pigments, like lapis lazuli or ochre, when possible.
The membrane prototype kit was being tested with real samples and the results were promising since one could get results with microsamples taken from those real artworks from the Getty Conservation Institute, National Gallery of Art (Washington DC) and Laboratório José de Figueiredo. Although the nanobiosensor had worked, the final NANOART kit to be commercialized is still to be decided on what to use: membrane or microfluidic chip, since the later is more sensitive and faster than the membrane used almost until the end of this project.
Dr. Ana Claro has opened up a new research line based on the combination of nanobiosensors and Cultural Heritage in the research group of Dr. Jesús Martínez de la Fuente (supervisor) at the Institute of Nanoscience of Aragón (INA) from University of Zaragoza. The NANOART research has used nanobiosensors based on Gold Nanoparticles (GNPs) to identify different proteinaceous binders’ nature present in microsamples of historical artworks.
This two-year project has trained Ana Claro in essential areas of microscopy, biochemistry and nanoparticle science; while at the same time offering her ample project management, supervisory and teaching experience. She introduced the Biotechnology applied to Cultural Heritage in Biotechnology degree through a final year project of a student, who decided to stay longer in the group and pursue his work on the same theme during his master (NANOMAT). Also another student with Engineer background took the opportunity to develop a new laser setup suitable to analyse the samples made over the membranes, one of the possible future NANOkits. She has presented the results of her research at four different scientific events during this two-year period: three international conferences and one national. She has established important collaborations as a result of her travel to collect samples and promote her work. Together with her collaborators she will pursue the effort to obtain joint research funding (Spanish, Portugal and E.U.) in the near future.
One important part of her research at The University of Zaragoza was her supervision of Masters project students along with her supervisor. In the academic year 2014/15 she supervised Javi Lou (Biotechnology Degree), while in 2015/16 she supervised Javi Lou and Alberto Blein (Nanomaterials Master). This has been a rewarding experience and both students are now thinking of their future Doctoral studies.
Another additional aspect of the fellowship was Dr Ana Claro collaboration with NanoImmunotech Company. Gave her a different perspective compared with the Academic Research, in the way the work and the timeline approaches are completely different. Giving television and radio interviews about the fellowship and getting the feeling that one is really getting in touch with the general public can be a very grateful experience.
Award of this Marie Curie Intra-European Fellowship has been an important step in Dr Ana Claro’s career and the knowledge gained in completely new research areas during this period has been crucial in determining her long-term development. Having gained the training, support and experience required to ideally position her as an independent researcher, Dr Claro feels that she is now capable to carry out internationally leading research.
Through the research of Medieval and Renaissance treatises, referring the manufacturing process of paints, such as: On Divers Arts (12th century), Il libro del art (14th century), The Strasburg Manuscript (late Middle Ages), The book on how to make colours (a late medieval Portuguese technical text on illumination), 5) Vasari on Technique (1550) and The B.M. Sloane 2052 (17th century), three classes of binders were identified: proteinaceous, glycosides and oil binders. Dr Ana Claro decided to start with the proteinaceous ones, meaning with animal glue, mainly composed by collagen and egg white.
After an exhaustive research two antibodies for collagen type I, and two for ovalbumin (mono and polyclonal), were selected, to further on build up the NANOKit.
Having the biomarkers selected, standard samples mimicking real samples were made (Figures 1 and 2) and the development of the protocol that would establish the way on how the samples should be treated previously to the detection was created.
Synthesis and functionalization of Gold Nanoparticles (GNPs) were fundamental steps since it provided the nanoparticles necessary for the nanobiosensor. Once synthesized, the GNPs were functionalized with the different targeting molecules, by following the functionalization methodologies developed by the NG2. These targeting molecules were covalently attached to the nanoparticles’ surface and in an oriented way, so they can successfully bind to their target molecules, giving rise to GNPs conjugates, also known as gold nanoprobes.
The methodology of the nanobiosensor used in this project has been developed by NG2: the photothermal nanobiosensor (Figure 3). This sensor is based on GNPs conjugates that generate heat upon being irradiated with a laser. This method is more sensitive than the colorimetric one (first one selected for the NANOART project) although requires a more complex mechanism to reveal the final result. This issue was solved with the development of a hard and software for the laser set up (Figures 4 and 5).
Analyzes performed with the model samples showed in most of the cases the expected results. Some exceptions occurred when an interaction (expected and reported in literature) between the pigments and the binder could happen, namely with minium, cinnabar and white lead. Knowing that one can recommend microsampling stable pigments, like lapis lazuli or ochre, when possible.
The membrane prototype kit was being tested with real samples and the results were promising since one could get results with microsamples taken from those real artworks from the Getty Conservation Institute, National Gallery of Art (Washington DC) and Laboratório José de Figueiredo. Although the nanobiosensor had worked, the final NANOART kit to be commercialized is still to be decided on what to use: membrane or microfluidic chip, since the later is more sensitive and faster than the membrane used almost until the end of this project.
Dr. Ana Claro has opened up a new research line based on the combination of nanobiosensors and Cultural Heritage in the research group of Dr. Jesús Martínez de la Fuente (supervisor) at the Institute of Nanoscience of Aragón (INA) from University of Zaragoza. The NANOART research has used nanobiosensors based on Gold Nanoparticles (GNPs) to identify different proteinaceous binders’ nature present in microsamples of historical artworks.
This two-year project has trained Ana Claro in essential areas of microscopy, biochemistry and nanoparticle science; while at the same time offering her ample project management, supervisory and teaching experience. She introduced the Biotechnology applied to Cultural Heritage in Biotechnology degree through a final year project of a student, who decided to stay longer in the group and pursue his work on the same theme during his master (NANOMAT). Also another student with Engineer background took the opportunity to develop a new laser setup suitable to analyse the samples made over the membranes, one of the possible future NANOkits. She has presented the results of her research at four different scientific events during this two-year period: three international conferences and one national. She has established important collaborations as a result of her travel to collect samples and promote her work. Together with her collaborators she will pursue the effort to obtain joint research funding (Spanish, Portugal and E.U.) in the near future.
One important part of her research at The University of Zaragoza was her supervision of Masters project students along with her supervisor. In the academic year 2014/15 she supervised Javi Lou (Biotechnology Degree), while in 2015/16 she supervised Javi Lou and Alberto Blein (Nanomaterials Master). This has been a rewarding experience and both students are now thinking of their future Doctoral studies.
Another additional aspect of the fellowship was Dr Ana Claro collaboration with NanoImmunotech Company. Gave her a different perspective compared with the Academic Research, in the way the work and the timeline approaches are completely different. Giving television and radio interviews about the fellowship and getting the feeling that one is really getting in touch with the general public can be a very grateful experience.
Award of this Marie Curie Intra-European Fellowship has been an important step in Dr Ana Claro’s career and the knowledge gained in completely new research areas during this period has been crucial in determining her long-term development. Having gained the training, support and experience required to ideally position her as an independent researcher, Dr Claro feels that she is now capable to carry out internationally leading research.
