Food safety, which is regarded as a global and expensive health problem, is often threatened with the entry of microorganisms, which reduce their storage time and cause acute illness. Another important global problem is the corrosion of metal and plastic structures, which is caused by various colonies of bacteria. The use of microorganisms that cause the biodegradation of materials is promising for solving the problem of environmental pollution with polymeric materials. That is, the problem of developing methods and markers for identifying microorganisms with high sensitivity and reproducibility is relevant. Thus, the actual task is a developing of methods of detection with high sensitivity, reproducibility.
The article presents the results of recognition and labeling of bacteria with a block-copolymer with fragments of oligonucleotide and fluoroalkyl alcohol, using luminescence spectroscopy and mass spectroscopy of secondary ions. Obtaining a hybrid block-copolymer with an oligonucleotide block was carried out in two stages. A fluorine-containing surface-active polymer with a terminal functional epoxy group was obtained at the first stage. At the second stage, an oligonucleotide which contains the primary amino group was bound with epoxy group of polymeric block. The first block was obtained by radical polymerization of N-vinylpyrrolidone (NVP). The process was initiated by the redox system of cerium salt - fluoroalkyl alcohol. An epoxy-containing cumine derivative was used as a chain transfer agent for control the colloid-chemical characteristics of the products and the introduction into their composition of the terminal functional epoxy fragment.
The effect of the length of hydrophobic fluoroalkyl and hydrophilic oligo(NVP) fragments on the size of micelles formed by oligomers in water was studied by dynamic light scattering. It was established that the size of oligomer particles below the CMC point increases with increasing a length of the fluoroalkyl fragment, which is not observed for concentrations above the CMC point. The observed effect is explained by the different ability of the fluoroalkyl fragment to be compacted inside the micellar core for different concentrations of oligomers in the solution.
The detection of bacteria in luminescent light and by secondary ion mass spectroscopy confirmed the possibility of using fluoride-based hybrid block-copolymer (NVP) with oligonucleotide block as a bacterial label.
1. Lazcka O., Campo F. J. D., Muñoz F. X., Pathogen detection: A perspective of traditional methods and biosensors, Biosensors and Bioelectronics, 22 (2007) 1205-1217.
https://doi.org/10.1016/j.bios.2006.06.036
2. Singh, S. Poshtiban, S. Evoy, Recent advances in bacteriophage based biosensors for foodborne pathogen detection, Sensors, 13 (2013) 1763- 1786.
https://doi.org/10.3390/s130201763
3. Bhardwaj, N., Bhardwaj, S. K., Nayak, M. K., Mehta, J., Kim, K. H., & Deep, A. (2017). Fluorescent nanobiosensors for the targeted detection of foodborne bacteria. TrAC Trends in Analytical Chemistry, 97, 120-135.
https://doi.org/10.1016/j.trac.2017.09.010
4. Schutt, E. G., Klein, D. H., Mattrey, R. M., & Riess, J. G. (2003). Injectable microbubbles as contrast agents for diagnostic ultrasound imaging: the key role of perfluorochemicals. Angewandte Chemie International Edition, 42(28), 3218-3235.
https://doi.org/10.1002/anie.200200550
5. Mao, Y., Akram, M., Shi, J., Wen, J., Yang, C., Jiang, J., & Tian, Y. (2019). Optical oxygen sensors based on microfibers formed from fluorinated copolymers. Sensors and Actuators B: Chemical, 282, 885-895.
https://doi.org/10.1016/j.snb.2018.11.143
6. Riess, J. G., & Krafft, M. P. (2006). Fluorocarbon emulsions as in vivo oxygen delivery systems: Background and chemistry. In Blood substitutes (p. 259-275). Academic Press.
https://doi.org/10.1016/B978-012759760-7/50033-0
7. Riess, J. G. (2005). Understanding the fundamentals of perfluorocarbons and perfluorocarbon emulsions relevant to in vivo oxygen delivery. Artificial cells, blood substitutes, and biotechnology, 33(1), 47-63.
https://doi.org/10.1081/BIO-200046659
8. Lehmler, H. J. (2007). Perfluorocarbon compounds as vehicles for pulmonary drug delivery. Expert opinion on drug delivery, 4(3), 247-262.
https://doi.org/10.1517/17425247.4.3.247
9. Porsch, C., Zhang, Y., Östlund, Å., Damberg, P., Ducani, C., Malmström, E., & Nyström, A. M. (2013). In Vitro Evaluation of Non-Protein Adsorbing Breast Cancer Theranostics Based on 19FPolymer Containing Nanoparticles. Particle & particle systems characterization, 30(4), 381-390.
https://doi.org/10.1002/ppsc.201300018
10. Xiong, S. D., Li, L., Jiang, J., Tong, L. P., Wu, S., Xu, Z. S., & Chu, P. K. (2010). Cationic fluorine-containing amphiphilic graft copolymers as DNA carriers. Biomaterials, 31(9), 2673-2685.
https://doi.org/10.1016/j.biomaterials.2009.12.014
11. Wang, M., Liu, H., Li, L., & Cheng, Y. (2014). A fluorinated dendrimer achieves excellent gene transfection efficacy at extremely low nitrogen to phosphorus ratios. Nature communications, 5, 3053.
https://doi.org/10.1038/ncomms4053
12. Liu, G., Fan, W., Li, L., Chu, P. K., Yeung, K. W., Wu, S., & Xu, Z. (2012). Novel anionic fluorine-containing amphiphilic self-assembly polymer micelles for potential application in protein drug carrier. Journal of Fluorine Chemistry, 141, 21-28.
https://doi.org/10.1016/j.jfluchem.2012.05.021
13. Krafft, M. P., & Riess, J. G. (2009). Chemistry, physical chemistry, and uses of molecular fluorocarbon-hydrocarbon diblocks, triblocks, and related compounds unique "apolar" components for self-assembled colloid and interface engineering. Chemical reviews, 109(5), 1714-1792.
https://doi.org/10.1021/cr800260k
14. Ameduri, B., & Vitale, A. (2014). Fluorinated Oligomers and Polymers in Photopolymerization. Chemical Reviews, 115, XY.
https://doi.org/10.1021/acs.chemrev.5b00120
15. Kinash, N. I., Paiuk, O. L., Dolynska, L. V., Nadashkevych, Z. Ya., & Hevus, O. I. (2017). Syntez novykh funktsionalnykh pokhidnykh kuminovoho spyrtu. Visnyk Nats. un-tu "Lvivska politekhnika". Seriia: Khimiia, tekhnolohiia rechovyn ta yikh zastosuvannia, (868), 40-44.
16. Oliveira, M., Andrade, G., Guerra, M., & Bernardo, F. (2003). Development of a fluorescent in situ hybridization protocol for the rapid detection and enumeration of Listeria monocytogenes in milk. Revista Portuguesa de Ciências Veterinárias, 119-124.