Диэлектрофоретическое осаждение белков на серебряные подложки для повышения эффективности спектрального анализа

Аleksandr М. МЕRZLIKIN; Il’ya А. RYZHIKOV; Ирина Анатольевна Богинская; Мarina V. SEDOVA; Andrey S. NABOKO; Victor I. POLOZOV; Sergey I. GUDKOV; Timur A. TERENT’EV

doi:10.24160/0013-5380-2026-3-4-13

Authors

Аleksandr М. МЕRZLIKIN
Il’ya А. RYZHIKOV
Ирина Анатольевна Богинская
Мarina V. SEDOVA
Andrey S. NABOKO
Victor I. POLOZOV
Sergey I. GUDKOV
Timur A. TERENT’EV

DOI:

https://doi.org/10.24160/0013-5380-2026-3-4-13

Keywords:

dielectrophoretic deposition, protein, surface-enhanced Raman spectroscopy

Abstract

The article suggests an approach that helps make spectral analysis more efficient. The proposed approach combines electrochemical sample preparation of a high-molecular analyte in the alternating electric field (AEF) mode followed by surface enhanced Raman spectroscopy (SERS). The method is of practical significance for the quantitative marker-free determination of clinically significant analytes. Human serum albumin (HSA) is used as a model analyte. The developed dielectrophoretic deposition method is aimed at improving the quality (the signal amplitude and the signal-to-noise ratio in the measured spectra) of the SERS spectra serving as an analytical signal. The range of detectable concentrations with AEF was found to be 0.01–10 g/L. The range of detectable concentrations for the AEF mode was an order of magnitude higher than it is for the constant electric field (CEF) mode. It is shown that precipitation is accompanied by redox reactions leading to the synthesis of silver nanoparticles (NPs) deposited in the form of Ag-HSA-Ag NPs sandwich structures. The self-assembled protein- and AgNP-based precipitate obtained after AEF treatment on a modified SERS substrate, from which the SERS spectra were measured, has demonstrated sufficient morphological homogeneity. This is confirmed by the standard deviation of the SERS spectral amplitude, which did not exceed 20% across the entire spectral range.

Author Biographies

Аleksandr М. МЕRZLIKIN

(Institute for Theoretical and Applied Electromagnetics RAS, Moscow, Russia) – Deputy Director for Science, Corresponding Member of RAS, Dr. Sci (Eng.).

Il’ya А. RYZHIKOV

(Institute for Theoretical and Applied Electromagnetics RAS, Moscow, Russia) – Head of Laboratory No. 4, Cand. Sci. (Eng.).

Ирина Анатольевна Богинская

(Institute for Theoretical and Applied Electromagnetics RAS, Moscow, Russia) – Leading Research Fellow, Cand. Sci. (Eng.).

Мarina V. SEDOVA

(Institute for Theoretical and Applied Electromagnetics RAS, Moscow, Russia) – Leading Research Fellow, Cand. Sci. (Eng.).

Andrey S. NABOKO

(Institute for Theoretical and Applied Electromagnetics RAS, Moscow, Russia) – Senior Research Fellow, Cand. Sci. (Eng.).

Victor I. POLOZOV

(Institute for Theoretical and Applied Electromagnetics RAS, Moscow, Russia) – Senior Research Fellow, Cand. Sci. (Eng.).

Sergey I. GUDKOV

(Institute for Theoretical and Applied Electromagnetics RAS, Moscow, Russia) – Research Fellow, Cand. Sci. (Eng.).

Timur A. TERENT’EV

(Institute for Theoretical and Applied Electromagnetics RAS, Moscow, Russia) – Research Engineer.

References

1. Blanch E.W. et al. Structural Characterization of Proteins and Viruses Using Raman Optical Activity. – Vibrational Spectroscopy, 2004, vol. 35, No. 1-2, pp. 87–92, DOI: 10.1016/j.vibspec.2003.12.005.

2. Rygula A. et al. Raman Spectroscopy of Proteins: A Review. – Journal of Raman Spectroscopy, 2013, vol. 44, No. 8, pp. 1061–1076, DOI: 10.1002/jrs.4335.

3. Keskin S., Çulha M. Label-Free Detection of Proteins from Dried-Suspended Droplets Using Surface Enhanced Raman Scattering. – Analyst, 2012, vol. 137, No. 11, pp. 2651–2657, DOI: 10.1039/c2an16296b.

4. Nemecek D., Stepanek J., Thomas Jr G.J. Raman Spectroscopy of Proteins and Nucleoproteins. – Current Protocols in Protein Science, 2013, vol. 71, No. 1, DOI: 10.1002/ 0471140864.ps1708s71.

5. Sharma B. et al. SERS: Materials, Applications, and the Future. – Materials Today, 2012, vol. 15, No. 1-2, pp. 16–25, DOI: 10.1016/S1369-7021(12)70017-2.

6. Fan M., Andrade G.F.S., Brolo A.G. A Review on the Fabrication of Substrates for Surface Enhanced Raman Spectroscopy and their Applications in Analytical Chemistry. – Analytica Chimica Acta, 2011, vol. 693, No. 1-2, pp. 7–25, DOI: 10.1016/j. aca.2011.03.002.

7. Das G. et al. Principal Component Analysis Based Methodology to Distinguish Protein SERS Spectra. – Journal of Molecular Structure, 2011, vol. 993, No. 1-3, pp. 500–505, DOI: 10.1016/j.mol-struc.2010.12.044.

8. Kahraman M., Wachsmann-Hogiu S. Label-Free and Direct Protein Detection on 3D Plasmonic Nanovoid Structures Using Surface-Enhanced Raman Scattering. – Analytica Chimica Acta, 2015, vol. 856, pp. 74–81, DOI: 10.1016/j.aca.2014.11.019.

9. Mitchell B.L. et al. Experimental and Statistical Analysis Methods for Peptide Detection Using Surface‐Enhanced Raman Spectroscopy. – Journal of Raman Spectroscopy, 2008, vol. 39, No. 3, pp. 380–388, DOI: 10.1002/jrs.1834.

10. Bhaskar S. et al. Single-Molecule Cholesterol Sensing by Integrating Silver Nanowire Propagating Plasmons and Graphene Oxide π-Plasmons on a Photonic Crystal-Coupled Emission Platform. – ACS Applied Optical Materials, 2022, vol. 1, No. 1, pp. 159–172, DOI: 10.1021/ acsaom.2c00026.

11. Boginskaya I.A. et al. Additional Enhancement of SERS Effect by a Surface Wave in Photonic Crystal. – Journal of Raman Spectroscopy, 2019, vol. 50, No. 10, pp. 1452–1461, DOI: 10.1002/ jrs.5680.

12. Boginskaya I.A. et al. Biological Object Determination by Raman Scattering Enhancement Supported on the Multilayer Dielectric Thin Film. – Progress in Electromagnetics Research Symposium-Spring (PIERS), 2017, pp. 3094–3097, DOI: 10.1109/PIERS.2017.8262287.

13. Stewart S., Fredericks P.M. Surface-Enhanced Raman Spectroscopy of Peptides and Proteins Adsorbed on an Electrochemically Prepared Silver Surface. – Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy, 1999, vol. 55, No. 7-8, pp. 1615–1640, DOI: 10.1016/S1386-1425(98)00293-5.

14. Grytsyk N. et al. Surface-Enhanced Resonance Raman Spectroscopy of Heme Proteins on a Gold Grid Electrode. – Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy, 2020, vol. 230, DOI: 10.1016/j.saa.2020.118081.

15. Vezvaie M., Brosseau C.L., Lipkowski J. Electrochemical Sers Study of a Biomimetic Membrane Supported at a Nanocavity Patterned Ag Electrode. – Electrochimica Acta, 2013, vol. 110, pp. 120–132, DOI: 10.1016/j.electacta.2013.03.139.

16. Fortunati S. et al. Rapid Quantification of SARS-CoV-2 Spike Protein Enhanced with a Machine Learning Technique Integrated in a Smart and Portable Immunosensor. – Biosensors, 2022, vol. 12, No. 6, DOI: 10.3390/bios12060426.

17. Ben Hassine A., Raouafi N., Moreira F.T.C. Novel Electrochemical Molecularly Imprinted Polymer-Based Biosensor for Tau Protein Detection. – Chemosensors, 2021, vol. 9, No. 9, DOI: 10.3390/chemosensors9090238.

18. Castano-Guerrero Y. et al. SERS and Electrochemical Impedance Spectroscopy Immunoassay for Carcinoembryonic Antigen. – Electrochimica Acta, 2021, vol. 366, DOI: 10.1016/j.electacta.2020.137377.

19. Walker K.A. et al. Proteomics Analysis of Plasma from Middle-Aged Adults Identifies Protein Markers of Dementia Risk in Later Life. – Science Translational Medicine, 2023, vol. 15, No. 705, DOI: 10.1126/scitranslmed.adf5681.

20. Moore C.M. et al. A Novel Microfluidic Dielectrophoresis Technology to Enable Rapid Diagnosis of Mycobacteria Tuberculosis in Clinical Samples. – The Journal of Molecular Diagnostics, 2023, vol. 25, No. 7, pp. 513–523, DOI: 10.1016/j.jmoldx.2023.04.005.

21. Sharma B., Sharma A. Microfluidics: Recent Advances Toward Lab‐On‐Chip Applications in Bioanalysis. – Advanced Engineering Materials, 2022, vol. 24, No. 2, DOI: 10.1002/adem.202100738.

22. Mehta M. et al. Laboratory‐on‐a‐Chip: A Multitasking De-vice. – Miniaturized Analytical Devices: Materials and Technology, 2022, pp. 91–103, DOI: 10.1002/9783527827213.ch5.

23. Boginskaya I. et al. Human Angiotensin I‐Converting Enzyme Study by Surface‐Enhanced Raman Spectroscopy. – Journal of Raman Spectroscopy, 2021, vol. 52, No. 9, pp. 1529–1539, DOI: 10.1002/jrs.6068.

24. Kurochkin I.N. et al. SERS for Bacteria, Viruses, and Protein Biosensing. – Macro, Micro, and Nano-Biosensors: Potential Applications and Possible Limitations, Cham: Springer International Publishing, 2021, pp. 75–94, DOI: 10.1007/978-3-030-55490-3_5.

25. Boginskaya I.A. et al. Additional Enhancement of Surface-Enhanced Raman Scattering Spectra of Myoglobin Precipitated Under Action of Laser Irradiation on Self-Assembled Nanostructured Surface of Ag Films. – Chemosensors, 2023, vol. 11, No. 6, DOI: 10.3390/chemosensors11060321.

26. Xia X. Bioinformatics and the Cell: Modern Computational Approaches in Genomics, Proteomics and Transcriptomics. Boston, MA: Springer US, 2007, 349 p., DOI: 10.1007/978-0-387-71337-3.

27. Podoynitsyn S.N. et al. Surface-Enhanced Raman Spectroscopy in Tandem with a Gradient Electric Field from 4-Mercaptophenylboronic Acid on Silver Nanoparticles. – Microchimica Acta, 2020, vol. 187, No. 10, DOI: 10.1007/s00604-020-04550-x.

28. Boginskaya I. et al. SERS-Active Substrates Nanoengineering Based on E-Beam Evaporated Self-Assembled Silver Films. – Applied Sciences, 2019, vol. 9, No. 19, DOI: 10.3390/app9193988.

29. Khaydarov R.A. et al. Electrochemical Method for the Synthesis of Silver Nanoparticles. – Journal of Nanoparticle Research, 2009, vol. 11, No. 5, pp. 1193–1200, DOI: 10.1007/s11051-008-9513-x.

30. Chiotelli E., Pilosio G., Le Meste M. Effect of Sodium Chloride on the Gelatinization of Starch: A Multimeasurement Study. – Biopolymers, 2002, vol. 63, No. 1, pp. 41–58, DOI: 10.1002/bip.1061.

31. Deegan R.D. et al. Contact Line Deposits in an Evaporating Drop. – Physical review E, 2000, vol. 62, No. 1, DOI: 10.1103/PhysRevE.62.756.

32. Synytsya A. et al. Raman Spectroscopic Study of Serum Albumins: An Effect of Proton‐ and γ‐Irradiation. – Journal of Raman Spectroscopy, 2007, vol. 38, No. 12, pp. 1646–1655, DOI: 10.1002/jrs.1884.

33. Jurasekova Z. et al. Spectroscopic and Molecular Modeling Studies on the Binding of the Flavonoid Luteolin and Human Serum Albumin. – Biopolymers, 2009, vol. 91, No. 11, pp. 917–927, DOI: 10.1002/ bip.21278.

34. Stewart S., Fredericks P.M. Surface-Enhanced Raman Spectroscopy of Amino Acids Adsorbed on an Electrochemically Prepared Silver Surface. – Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy, 1999, vol. 55, No. 7-8, pp. 1641–1660, DOI: 10.1016/S1386- 1425(98)00294-7.

35. Yang S. et al. Electrochemically Created Highly Surface Roughened Ag Nanoplate Arrays for SERS Biosensing Applications. – Journal of Materials Chemistry C, 2014, vol. 2, No. 39, pp. 8350–8356, DOI: 10.1039/c4tc01276c.

36. Zhou H., Kneipp J. Electrodeposition of Silver Nanostructures in Ethanol for Sensitive Electrochemical SERS Detection. – ACS Applied Nano Materials, 2024, vol. 7, No. 1, pp. 1300–1309, DOI: 10.1021/acsanm.3c05322.

37. Oriňáková R. et al. Electrochemical Deposition of SERS Active Nanostructured Silver Films. – International Journal of Electrochemical Science, 2013, vol. 8, No. 1, pp. 80–99, DOI: 10.1016/s1452-3981(23)14004-1.

38. Kim S. et al. Early and Direct Detection of Bacterial Signaling Molecules through One-Pot Au Electrodeposition onto Paper-Based 3D SERS Substrates. – Sensors and Actuators B: Chemical, 2022, vol. 358, DOI: 10.1016/j.snb.2022.131504.

39. Zhan Y. et al. Facile Electrochemical Surface-Alloying and Etching of Au Wires to Enable High-Performance Substrates for Surface Enhanced Raman Scattering. – Nano Materials Science, 2024, vol. 6, No. 3, pp. 305–311, DOI: 10.1016/j.nanoms.2023.05.002.

40. Chung C.K., Yu C.Y. Unique High-Performance Metal-Nanoparticle-Free SERS Substrate with Rapid-Fabricated Hybrid 3D-Nano-Micro-Cavities Anodic Alumina for Label-Free Detection. – Applied Surface Science, 2023, vol. 635, DOI: 10.1016/j. apsusc.2023.157731.

41. Nechaeva N.L. et al. Multiscale Flaked Silver SERS-Substrate for Glycated Human Albumin Biosensing. – Analytica Chimica Acta, 2020, vol. 1100, pp. 250–257, DOI: 10.1016/j.aca.2019.11.072.

42. Rai A. et al. Hottest Hotspots from the Coldest Cold: Welcome to Nano 4.0. – ACS Applied Nano Materials, 2022, vol. 5, No. 9, pp. 12245–12264, DOI: 10.1021/acsanm.2c02556.

43. Akin M.S. et al. Large Area Uniform Deposition of Silver Nanoparticles through Bio-Inspired Polydopamine Coating on Silicon Nanowire Arrays for Practical SERS Applications. – Journal of Materials Chemistry B, 2014, vol. 2, No. 30, pp. 4894–4900, DOI: 10.1039/c4tb00616j.

44. Pérez-Jiménez A.I. et al. Surface-Enhanced Raman Spectroscopy: Benefits, Trade-Offs and Future Developments. – Chemical Science, 2020, vol. 11, No. 18, pp. 4563–4577, DOI: 10.1039/d0sc00809e.

45. Azimi S., Docoslis A. LESS is More: Achieving Sensitive Protein Detection by Combining Electric Field Effects and Surface-Enhanced Raman Scattering. – Sensors and Actuators B: Chemical, 2023, vol. 393, DOI: 10.1016/j.snb.2023.134250.

---

Работа выполнена при поддержке Министерства науки и высшего образования РФ в рамках Государственного задания FFUR-2024-0010