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Study of the reusability and stability of nylon nanofibres as an antibody immobilisation surface
(Beilsein Institut, 2024-01-15) Peraile, Inés; Gil García, Matilde; González López, Laura, Laura; Dabbagh Escalante, Nushin Alba; Cabria Ramos, Juan Carlos; Lorenzo Lozano, Paloma; Ministerio de Economía y Competitividad (MINECO)
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.
Biofunctionalization of nylon nanofibers to be used in immunobiosensor for biological warfare agents detecting
(Formatex Research Center, 2018) Peraile, Inés; Lorenzo Lozano, Paloma; Murillo, N.; Maudes, J.; Rozas Sanz, Gabriel; Pérez Márquez, Ana; González López, Laura; Cabria Ramos, Juan Carlos; Gil García, Matilde
The use of biological warfare agents involves a growing threat to society. Thus, the most countries have been forced to increase resources in research for their detection and identification. One of the critical points in the effective fight against these agents is the development of devices that allow their detection and early identification. The best choice is the immunobiosensors easy to use on-site. However, how the antibody is attached to the biosensor surface, in terms of density and orientation, will determine the diagnosis capability of the device. In this study, both a functional nanofiber able to increase the surface / volume relation, and a chemicallysimilar planar membrane were used as support for the immobilization of antibodies. Different antibody immobilization systems were carried out to biofunctionalize both surfaces: passive adsorption, covalent bond by glutaraldehyde and well-oriented immobilization by protein A/G. Our results showed that nanofibers in combination with protein A/G were a very effective immunocapture system for being used in a biosensor.
lnactivation and identification of Bacillus anthracis spores
(Wageningen Academic Publishers, 2013-12) Gil García, Matilde; Peraile, Inés; Jiménez Pérez, M. V.; Cabria Ramos, Juan Carlos; Lorenzo Lozano, Paloma
Bacillus anthracis endospores must be handled ata biosafety level-3 (BSL-3} eontainment facility, therefore an appropriate inaetivation procedure is essential in order to both detect thcm in a reliable and rapid way and to avoid human infections from antrax-contamined samples if a biosafety level-3 laboratory is not availablc. Unfortunatcly, donnant spores exhibit incredible hardiness against gennicidal agents. The most effective methods for inactivating spores are usually the chemical treatment, heat inactivation (boiling, moist hcat, dry heat) and irTadiation (microwave, UV, gamma, electron beam). The inactivation method used must alter thc cell structure as little as possible in order to ensure a corree! diagnosis by diferent techniques. Several kinds ofphysical and chemical inactivation methods wcrc assessed in order to define the conditions that ensure inactivation of endospores of B. anthracis 34F2 Sterné and pcrmit, at the same time, its identification by PCR and immunoassays techniques.
