Determination of Protein Amount in Nanosized Synthetic Liposomes by Surface Effect Raman Spectroscopy (SERS)

Abstract

Accurate characterization of synthetic liposomes is essential since they give information about the vesicu-lar structures in bodily fluids such as extracellular vesicles. The characterization tasks are generally the determination of the sizes of the liposomes and the profiling of the liposomes' content. Optical tweezers and Surface Enhanced Raman Spectroscopy (SERS) were used to profile the nanosized liposomes. The size distribution of the trapped liposomes (140 nm on average) was found by using Einstein's Brownian motion equation, consistent with the size distribution obtained from dynamic light scattering measure-ments. Besides, Gramicidin-encapsulated liposomes were measured using SERS, and statistically signifi-cant differentiation was found in Raman intensities between liposome populations with altering concentra-tions of proteins. This study uniquely measured size distributions of nano-sized liposomes with conven-tional optical tweezers (without plasmonics) and determined the chemical differences between empty and protein encapsulated liposomes with high accuracy using Raman spectroscopy

Keywords

• Liposome
• optical trapping
• surface enhanced Raman spectroscopy
• Gramicidin
• protein determination

Supporting Institution

İstinye Üniversitesi

Thanks

The author thanks Boğazici University BUMILAB for their permission to perform the measurements in their facilities. The author thanks Şebnem Seherler for their help in the preparation of the liposomes.

References

1. Akbarzadeh, A., Sadabady, R.R., Davaran, S., Joo, S. W., Zarghami, N., Hanifehpour, Y., Samiei, M., Kouhi, M., Nejati-Koshk, K., (2013). Liposome: classification, preparation, and applications. Nanoscale research letters, 8(1), 1-9. Retrieved from: https://doi.org/10.1186/1556-276X-8-102
2. Alavi, M., Karimi, N., Safaei, M., (2017). Application of various types of liposomes in drug delivery systems. Advanced pharmaceutical bulletin, 7(1), 3-9, Retrieved from: https://doi.org/ 10.15171/apb.2017.002
3. Allen, T.M., Cullis, P.R, (2013). Liposomal drug delivery systems: from concept to clinical applications. Advanced drug delivery reviews,65(1), 36-48. Retrieved from: https://doi.org/10.1016/j.addr.2012.09.037
4. Ashkin, A., (1997). Optical trapping and manipulation of neutral particles using lasers. Proceedings of the National Academy of Sciences, 94(10), 4853-4860. Retrieved from: https://doi.org/10.1073/pnas.94.10.4853
5. Bruzas, I., Brinson, B. E., Gorunmez, Z., Lum, W., Ringe, E., & Sagle, L. (2019). Surface-enhanced Raman spectroscopy of fluid-supported lipid bilayers. ACS applied materials & interfaces, 11(36), 33442-33451. Journal of Advanced Research in Natural and Applied Sciences 2023, Cilt 9, Sayı 4, Sayfa: 912-922 920
6. Chan, J.W., Winhold, H., Lane, S.M., Huser, T., (2005). Optical trapping and coherent anti-Stokes Raman scattering (CARS) spectroscopy of submicron-size particles. IEEE Journal of selected topics in quantum electronics, 11(4), 858-863. Retrieved from: https://doi.org/10.1109/JSTQE.2005.857381
7. Chaney, S.B., Shanmukh, S., Dluhy, R.A., Zhao, Y.P., (2005). Aligned silver nanorod arrays produce high sensitivity surface-enhanced Raman spectroscopy substrates. Applied Physics Letters, 87(3), 031908. Retrieved from: https://doi.org/10.1063/1.1988980
8. Cheng, J.-X., Jia, Y.K., Zheng, G., Xie, X. S., (2002). Laser-scanning coherent anti-Stokes Raman scattering microscopy and applications to cell biology. Biophysical journal, 83(1), 502-509. Retrieved from: https://doi.org/10.1016/S0006-3495(02)75186-2

9. Cherney, D.P., Conboy, J.C., Harris, J.M., (2003). Optical-trapping Raman microscopy detection of single unilamellar lipid vesicles. Analytical chemistry, 75(23),6621-6628. Retrieved from:https://doi.org/10.1021/ac034838r
10. Cherney, D.P., Bridges, T.E., Harris, J.M., (2004). Optical trapping of unilamellar phospholipid vesicles: investigation of the effect of optical forces on the lipid membrane shape by confocal-Raman microscopy. Analytical chemistry, 76(17), 4920-4928. Retrieved from: https://doi.org/10.1021/ac0492620
11. Dufresne, E.R., Corwin, E. I., Greenblatt, N. A., Ashmore, J.,Wang, D. Y. , Dinsmore, A. D., Cheng, J. X., Xie, X. S., Hutchinson, J. W., Weitz, D. A.,(2003). Flow and fracture in drying nanoparticle suspensions. Physical review letters, 91(22), 224501. Retrieved from: https://doi.org/10.1103/PhysRevLett.91.224501
12. Evans, C.L., Potma, E.O., Puoris’haag, M., Xie, X.S., (2005). Chemical imaging of tissue in vivo with video-rate coherent anti-Stokes Raman scattering microscopy. Proceedings of the national academy of sciences, 102(46),16807-16812. Retrieved from: https://doi.org/10.1073/pnas.0508282102