Izvestiya of Saratov University.

Physics

ISSN 1817-3020 (Print)
ISSN 2542-193X (Online)


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Larionova O. S., Drevko Y. B., Khanadeev V. A., Gorshunova S. V., Kozlov E. S., Larionov S. V. Analysis of protein fractions of water-soluble peptides by dynamic light scattering. Izvestiya of Saratov University. Physics , 2023, vol. 23, iss. 1, pp. 37-45. DOI: 10.18500/1817-3020-2023-23-1-37-45, EDN: DLHAFH

This is an open access article distributed under the terms of Creative Commons Attribution 4.0 International License (CC-BY 4.0).
Published online: 
01.03.2023
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(downloads: 216)
Language: 
Russian
Article type: 
Article
UDC: 
60:543.645.6:535.375.55
EDN: 
DLHAFH

Analysis of protein fractions of water-soluble peptides by dynamic light scattering

Autors: 
Larionova Olga Sergeevna, Federal State Budgetary Educational University of Higher Education “Saratov State University of genetics, biotechnology and engineering named after N. I. Vavilov”
Drevko Yaroslav Borisovech, Federal State Budgetary Educational University of Higher Education “Saratov State University of genetics, biotechnology and engineering named after N. I. Vavilov”
Khanadeev Vitaly Andreevich, Institute of Biochemistry and Physiology of Plants and Microorganisms, Saratov Scientific Centre of the Russian Academy of Sciences (IBPPM RAS)
Gorshunova Sofia Vladimirovna, Federal State Budgetary Educational University of Higher Education “Saratov State University of genetics, biotechnology and engineering named after N. I. Vavilov”
Kozlov Evgeny Sergeevich, Federal State Budgetary Educational University of Higher Education “Saratov State University of genetics, biotechnology and engineering named after N. I. Vavilov”
Larionov Sergey Vasilievich, Federal State Budgetary Educational University of Higher Education “Saratov State University of genetics, biotechnology and engineering named after N. I. Vavilov”
Abstract: 

Background and Objectives: Currently, antimicrobial peptides are one of the main sources of alternative antibiotics because they can easily interact with bacterial peptidoglycan by penetrating or dissolving biofilms with minimal side effects. However, from a practical point of view, questions remain open about choosing the optimal method for obtaining and scaling the process of isolating peptides from insect biomass, as well as analyzing drug prototypes. In this regard, the search for methods of analysis and control of protein fractions of water-soluble peptides used for the subsequent development of antibacterial drugs based on them is an urgent task. The aim of this work was to study the protein fractions of water-soluble peptides isolated from Musca domestica larvae by dynamic light scattering. Materials and Methods: The results of the study of fractions of water-soluble peptides from Musca domestica larvae by dynamic light scattering are presented. The selection of optimal methods for the analysis and control of prototypes of antibacterial drugs based on antimicrobial peptides will reduce the time of research and ensure the accuracy of the results obtained. Results: It has been found that all the analyzed peptides have a sufficiently high stability in the aqueous medium which is confirmed by values of the zeta potential from −11.2 mV to −12 mV. The peptides at a concentration of 666 µg/ml with a molecular weight of less than 3.5 kDa, it has been found that their size was in the range of 68–142 nm; with a molecular weight of 3.5–7 kDa – 43–68 nm; with a molecular weight of 7–14 kDa – 43–105 nm; with a molecular weight of more than 14 kDa – 79–190 nm. The use of the dynamic light scattering method for the control and analysis of protein fractions of water-soluble peptides is established. The use of this method will reduce the time of analysis, identify micro-impurities, ensure simplicity of execution and almost complete absence of consumables. Conclusion: Use of dynamic light scattering is justified as a fast method of analyzing the obtained fractions of peptides, establishing the absence of trace impurities. Taking into account the almost complete absence of consumables, reduced analysis time and ease of execution in comparison with high-performance liquid chromatography, this detection method can be successfully used in everyday practice. 

Acknowledgments: 
The research was carried out at the expense of the grant of the Russian Science Foundation No. 22-26-00167, https://rscf.ru/project/22-26-00167/
Reference: 
  1. Valdez-Miramontes C. E., De Haro-Acosta J., Aréchiga-Flores C. F., Verdiguel-Fernández L., Rivas-Santiago B. Antimicrobial peptides in domestic animals and their applications in veterinary medicine. Peptides, 2021, vol. 142, article no. 170576. https://doi.org/10.1016/j.peptides.2021.170576
  2. Aamra H., Khan Farooq-Ahmad, Jahan H., Zafar M., Ali H., Farzana S. Synthesis of novel benzimidazole containing antimicrobial peptides (AMPs) with significant inhibitory effect on multidrug resistant strain of Salmonella typhimurium. Synth. Comm., 2021, vol. 51, no. 23, pp. 3620–3628. https://doi.org/10.1080/00397911.2021.1986841
  3. Berne B. J., Pecora R. Dynamic Light Scattering: With Applications to Chemistry, Biology, and Physics. USA, Dover Publ., Mineola, N. Y., 2000. 384 p. https://doi.org/10.1021/ed054pA430.1
  4. Pecora R. Dynamic Light Scattering – applications of Photon Crrelation Spectroscopy. Plenum Press, 1985. 420 p. https://doi.org/10.1002/bbpc.19870910455
  5. Attri A. K., Minton A. P. New methods for measuring macromolecular interactions in solution via static light scattering: Basic methodology and application to nonassociating and self-associating proteins. Anal. Biochem. : Meth. Biol. Sci., 2005, vol. 337, pp. 103–110. https://doi.org/10.1016/j.ab.2004.09.045
  6. Thurston G. M. Liquid-liquid phase separation and static light scattering of concentrated ternary mixtures of bovine alpha and gammaB crystallins. J. Chem. Phys., 2006, vol. 124, article no. 134909. https://doi.org/10.1063/1.2168451
  7. António M., Lima T., Vitorino R. L., Daniel-da-Silva A. Label-free dynamic light scattering assay for C-reactive protein detection using magnetic nanoparticles. Anal. Chim. Acta, 2022, vol. 1222, no. 9, article no. 340169. https://doi.org/10.1016/j.aca.2022.340169
  8. Holloway L., Roche A., Marzouk S., Uddin S., Ke P., Ekizoglou S., Curtis R. Determination of Protein-Protein Interactions at High Co-Solvent Concentrations Using Static and Dynamic Light Scattering. J. Pharm. Sci., 2020, vol. 109, no. 9, pp. 2699–2709. https://doi.org/10.1016/j.xphs.2020.05.023
  9. Vasil’eva I. A., Anarbaev R. O., Moor N. A., Lavrik O. I. Dynamic light scattering study of base excision DNA repair proteins and their complexes. Biochim. Biophy. Acta (BBA) – Prot. Proteom., 2019, vol. 1867, no. 3, pp. 297–305. https://doi.org/10.1016/j.bbapap.2018.10.009
  10. Fukushima K., Okada A., Sasaki K., Kishimoto S., Fukushima S., Hamori M., Nishimura A., Shibata N., Shirai T., Terauchi R., Kubo T., Sugioka N. Population Pharmacokinetic–Toxicodynamic Modeling and Simulation of Cisplatin-Induced Acute Renal Injury in Rats: Effect of Dosing Rate on Nephrotoxicity. J. of Pharm. Sci., 2016, vol. 105, no. 1, pp. 324–332. https://doi.org/10.1016/j.xphs.2015.10.022
  11. Meyer W. V., Smart A. E., Wegdam G. H., Brown R. G. W. Photon correlation and scattering: Introduction to the feature issue. Appl. Opt., 2006, vol. 45, pp. 2149–2154. https://doi.org/10.1364/AO.45.002149
  12. Mahatnirunkul T., Tomlinson D. C., McPherson M. J., Millnera P. A. One-step gold nanoparticle size-shift assay using synthetic binding proteins and dynamic light scattering. Sensors and Actuators B : Chemical, 2022, vol. 361, article no. 131709. https://doi.org/10.1016/j.snb.2022.131709
  13. Khan S. A., Degrasse J. A., Yakes B. J., Croley T. R. Rapid and sensitive detection of cholera toxin using gold nanoparticle-based simple colorimetric and dynamic light scattering assay. Anal. Chim. Acta, 2015, vol. 892, pp. 167–174. https://doi.org/10.1016/j.aca.2015.08.029
  14. Miao X., Ling L., Shuai X. Sensitive detection of glucose in human serum with oligonucleotide modified gold nanoparticles by using dynamic light scattering technique. Biosens. Bioelectron., 2013, vol. 41, pp. 880–883. https://doi.org/10.1016/j.bios.2012.09.015
  15. Li-na M. A., Dian-jun L. I. U., Zhen-xin W. Gold nanoparticle-based dynamic light scattering assay for mercury ion detection. Chin. J. Anal. Chem., 2014, vol. 42, iss. 3, pp. 332–336.
  16. Alami A. El., Lagarde F., Huo Q., Zheng T., Baitoul M., Daniel P. Acetylcholine and acetylcholinesterase inhibitors detection using gold nanoparticles coupled with dynamic light scattering. Sensors Int., 2020, vol. 1, article no. 100007. https://doi.org/10.1016/j.sintl.2020.100007
  17. Zheng X. T., Goh W. L., Yeow P., Lane D. P., Ghadessy F. J., Tan Y. N. Ultrasensitive dynamic light scattering based nanobiosensor for rapid anticancer drug screening. Sensor. Actuator. B Chem., 2019, vol. 279, pp. 79–86. https://doi.org/10.1016/j.snb.2018.09.088
  18. Levin A. D., Ringaci A., Alenichev M. K., Drozhzhennikova E. B., Shevchenko K. G., Cherkasov V. R., Nikitin M. P., Nikitin P. I. Dynamic light scattering biosensing based on analyte-induced inhibition of nanoparticle aggregation. Anal. Bioanal. Chem., 2020, vol. 412, pp. 3423–3431. https://doi.org/10.1007/s00216-020-02605-9
  19. Levin A. D., Filimonov I. S., Alenichev M. K., Goidina T. A. Mathematical modeling of nanosensor systems based on dynamic light scattering. Nano Technol. Russ., 2018, vol. 13, pp. 406–413. https://doi.org/10.1134/S1995078018040092
  20. Sergeeva I. A., Khitrina K. A., Krot A. R., Sukneva A. V., Petrova G. P. Investigation of the Interaction and Dynamics of Collagen and Collagenase Molecules in Solutions by Dynamic Light Scattering. Izvestiya of Saratov University. Physics, 2017, vol. 17, iss. 3, pp. 171–178 (in Russian). https://doi.org/10.18500/1817-3020-2017-17-3-171-178
  21. Witten K. G., Bretschneider J. C., Eckert T., Richtering W., Simon U. Assembly of DNA-functionalized gold nanoparticles studied by UV/Vis-spectroscopy and dynamic light scattering. Phys. Chem. Chem. Phys., 2008, vol. 10, pp. 1870–1875. https://doi.org/10.1039/b719762d
  22. Bhattacharjee S. DLS and zeta potential – What they are and what they are not? J. Contr. Release, 2016, vol. 235, pp. 337–351. https://doi.org/10.1016/j.jconrel.2016.06.017
  23. Krylova L. S., Drevko B. I., Faust E. A., Remizov E. K., Smirnova K. Yu., Drevko Ya. B., Borodina M. A., Osina T. S., Larionova O. S. Saratov State Vavilov Agrarian University. Composition of antimicrobial peptides obtained from Musca domestica larvae and method of its preparation. Patent RF no. RU 2714128 C1 (in Russian).
  24. Van der Zande B. M. I., Dhont Jan K. G., Bohmer Marcel R., Philipse A. P. Colloidal dispersions of gold rods characterized by dynamic light scattering and electrophoresis. Langmuir, 2000, vol. 16, pp. 459–464. https://doi.org/10.1021/la990043x
  25. Liu X., Huo Q. A washing-free and amplifi cation-free one-step homogeneous assay for protein detection using gold nanoparticle probes and dynamic light scattering. J. Immunol. Meth., 2009, vol. 349, pp. 38–44. https://doi.org/10.1016/j.jim.2009.07.015
  26. Khlebtsov B. N., Pylaev T. E., Khanadeev V. A., Khlebtsov N. G. Application of Dynamic Light Scattering and Absorption Spectroscopy to Studies of Systems with Colloidal Gold Nanoparticles + DNA. Izvestiya of Saratov University. Physics, 2017, vol. 17, iss. 3, pp. 136–149 (in Russian). https://doi.org/10.18500/1817-3020-2017-17-3-136-149 
Received: 
30.08.2022
Accepted: 
05.10.2022
Published: 
01.03.2023