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Analysis of human blood plasma and hen egg white by chiroptical spectroscopic methods (ECD, VCD, ROA)

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Abstract

Chiroptical methods are widely used in structural and conformational analyses of biopolymers. The application of these methods to investigations of biofluids would provide new avenues for the molecular diagnosis of protein-misfolding diseases. In this work, samples of human blood plasma and hen egg white were analyzed using a combination of conventional and chiroptical methods: ultraviolet absorption/electronic circular dichroism (UV/ECD), Fourier transform infrared absorption/vibrational circular dichroism (FTIR/VCD), and Raman scattering/Raman optical activity (Raman/ROA). For comparison, the main components of these substances—human serum albumin (HSA) and ovalbumin (Ova)—were also analyzed by these methods. The ultraviolet region of the ECD spectrum was analyzed using the CDNN CD software package to evaluate the secondary structures of the proteins. The UV/ECD, FTIR/VCD, and Raman/ROA spectra of the substances were quite similar to those of the corresponding major proteins, while some differences were also detected and explained. The conclusions drawn from the FTIR/VCD and Raman/ROA data were in good agreement with the secondary structures calculated from ECD. The results obtained in this work demonstrate that the chiroptical methods used here can be applied to analyze not only pure protein solutions but also more complex systems, such as biological fluids.

Analysis of human blood plasma and hen egg white by ECD, VCD and ROA.

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Acknowledgements

This work was supported by the Ministry of Education, Youth, and Sports of the Czech Republic (project No. CEZ: MSM6046137307), the Ministry of Health of the Czech Republic (project No. NT13259), the Complex Oncological Program (project PRVOUK P27), the Czech Science Foundation (project No. P208/11/0105), and Specific University Research MSMT (project No. 21/2012–A1_FCHI_2012_003, A2_FCHI_2012_015, and A2_FCHI_2012_045). The Jasco J-815 spectropolarimemeter and the ChiralRAMAN-2XTM instrument (BioTools, Inc.) were obtained through the financial support of the Prague Operational Program for Competitiveness (project No. CZ.2.16/3.100/22197).

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  1. Department of Analytical Chemistry, Institute of Chemical Technology Prague, Technická 5, 166 28, Prague 6, Czech Republic

    Alla Synytsya, Miluše Judexová, Tomáš Hrubý, Michal Tatarkovič & Vladimír Setnička

  2. Department of Oncology, First Faculty of Medicine, Charles University in Prague and General University Hospital in Prague, Ke Karlovu 11, 120 00, Prague 2, Czech Republic

    Michaela Miškovičová & Luboš Petruželka

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  1. Alla Synytsya
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Correspondence to Alla Synytsya.

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Synytsya, A., Judexová, M., Hrubý, T. et al. Analysis of human blood plasma and hen egg white by chiroptical spectroscopic methods (ECD, VCD, ROA). Anal Bioanal Chem 405, 5441–5453 (2013). https://doi.org/10.1007/s00216-013-6946-6

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