© 2024 Peraile et al.; licensee Beilstein-Institut.Peraile, InésGil García, MatildeGonzález López, Laura, LauraDabbagh Escalante, Nushin AlbaCabria Ramos, Juan CarlosLorenzo Lozano, Paloma2026-09-092026-09-092024-01-15Beilstein Journal of Nanotechnology 15: 83-94https://www.beilstein-journals.org/bjnano/articles/15/8https://hdl.handle.net/20.500.12666/1891Biological Defence Area, Department of NBC Defence Systems and Energetic Materials, National Institute for Aerospace Technology “Esteban Terradas” (INTA)-Campus La Marañosa, Ctra. M-301, Km 10, 28330, San Martín de la Vega, Madrid, Spain. This article is part of the thematic issue "Nanomaterials for biomedical applications".We would like to thank the whole team at Nieves Murillo of TECNALIA – Industry and Transport Division for providing the nylon NFs. (1) The samples must be dissolved in buffer with a physiological pH value before testing them. (2) The sensing method used was fluorescence as it is a simple method that does not require any additional steps for its determination. However, even though the aim of the study was to evaluate the reusability and stability of NFs, the sensitivity of the system could be improved by using another more accurate sensing system. (3) Because of the pore size of the nanofibres, they cannot be used for the detection of bacteria, rickettsiae, or fungi (i.e., they cannot be used for the detection of prokaryotic or eukaryotic cells). They could, therefore, be used for the determination of exogenously produced biotoxins and virulence factors, as well as for the detection of viruses and biomarkers in clinical samples (e.g., hormones and biomolecules). (4) The data could be generalised not only for the measurement of warfare agents, but also for the diagnosis of water and food contamination and for the clinical diagnosis of infectious agents and biomarkers.In the case of a biological threat, early, rapid, and specific detection is critical. In addition, ease of handling, use in the field, and low-cost production are important considerations. Immunological devices are able to respond to these needs. In the design of these immunological devices, surface antibody immobilisation is crucial. Nylon nanofibres have been described as a very good option because they allow for an increase in the surface-to-volume ratio, leading to an increase in immunocapture efficiency. In this paper, we want to deepen the study of other key points, such as the reuse and stability of these nanofibres, in order to assess their profitability. On the one hand, the reusability of nanofibres has been studied using different stripping treatments at different pH values on the nylon nanofibres with well-oriented antibodies anchored by protein A/G. Our study shows that stripping with glycine buffer pH 2.5 allows the nanofibres to be reused as long as protein A/G has been previously anchored, leaving both nanofibre and protein A/G unchanged. On the other hand, we investigated the stability of the nylon nanofibres. To achieve this, we analysed any loss of immunocapture ability of well-oriented antibodies anchored both to the nylon nanofibres and to a specialised surface with high protein binding capacity. The nanofibre immunocapture system maintained an unchanged immunocapture ability for a longer time than the specialised planar surface. In conclusion, nylon nanofibres seem to be a very good choice as an antibody immobilisation surface, offering not only higher immunocapture efficiency, but also more cost efficiency as they are reusable and stable.engAttribution 4.0 Internationalhttp://creativecommons.org/licenses/by/4.0/BiosensorBiothreatImmunodetectionNanofibreNyloninfo:eu-repo/semantics/articlehttps://doi.org/10.3762/bjnano.15.8http://dx.doi.org/10.13039/501100003329info:eu-repo/semantics/openAccess