4.7 Article

Single-step purified R-phycoerythrin transmits cellular imaging functionalities in vitro

Journal

INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
Volume 194, Issue -, Pages 563-570

Publisher

ELSEVIER
DOI: 10.1016/j.ijbiomac.2021.11.099

Keywords

Gracilaria corticata; Phycobiliprotein; R-phycoerythrin; SDS-PAGE; Physico-chemical properties; In vitro cell imaging

Funding

  1. National Natural Science Foundation of China [21476135]
  2. Educational Commission of Guangdong Province, China [2016KZDXM014]
  3. National Natural Science Foundation of Guangdong Province, China [2017A030307014]
  4. National Research Foundation of Korea (NRF) [2021R1I1A1A01046207]
  5. National Research Foundation of Korea [2021R1I1A1A01046207] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)

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The purification of R-phycoerythrin from Gracilaria corticata was efficiently achieved using a single-step chromatographic method based on polyacrylamide gel electrophoresis, maintaining structural integrity, UV-Vis spectrum characteristics, and stability. In vitro studies demonstrated the effective cell imaging properties of purified R-PE without impacting cell proliferation in Vero and Hep-2 cell lines.
A single-step and rapid chromatographic method-based purification of Gracilaria corticata (J. Agardh) R-phycoerythrin (R-PE) was attained using polyacrylamide gel electrophoresis (PAGE) technique without affecting structural integrity. The purified R-PE had a characteristic UV-Vis spectrum with three absorbance maxima at 496, 535, and 565 nm, and fluorescence at 575 nm. R-PE was obtained with a purity index of 4.2 and a recovery yield of 44.3%. SIDS-PAGE analysis exhibited three sub-units i.e., 18, 21, and 31 kDa, which corresponds to alpha, beta, and gamma, respectively. This report's purification process was considered less time-consuming and could be efficiently applied to purify phycobiliproteins. The purified R-PE showed optimal stability up to 6 h at pH 7.0 when exposed to light (3000 lx), while the temperature at which the maximum stability was retained was at 20 degrees C. The cellular imaging property of R-PE was effectively implemented to evaluate its credentials without affecting the cell proliferation of Vero and Hep-2 cell lines with the higher IC50 concentrations in vitro. Under fluorescence microscopy and flow cytometry analysis, purified R-PE displayed the characteristic affinity towards cell imaging functions in preliminary in vitro studies.

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