TY - JOUR
T1 - Single-step purified R-phycoerythrin transmits cellular imaging functionalities in vitro
AU - Sathuvan, Malairaj
AU - Thangam, Ramar
AU - Venkateshbabu, Gopal
AU - Cheong, Kit Leong
AU - Kang, Heemin
AU - Liu, Yang
N1 - Funding Information:
The authors are thankful to the National Natural Science Foundation of China ( 21476135 ), the Educational Commission of Guangdong Province , China ( 2016KZDXM014 ), and the National Natural Science Foundation of Guangdong Province , China ( 2017A030307014 ) for the financial support. This work is also supported by the National Research Foundation of Korea (NRF) grant ( 2021R1I1A1A01046207 ).
Publisher Copyright:
© 2021 Elsevier B.V.
PY - 2022/1/1
Y1 - 2022/1/1
N2 - 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%. SDS-PAGE analysis exhibited three sub-units i.e., 18, 21, and 31 kDa, which corresponds to α, β, and γ, 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 °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.
AB - 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%. SDS-PAGE analysis exhibited three sub-units i.e., 18, 21, and 31 kDa, which corresponds to α, β, and γ, 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 °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.
KW - Gracilaria corticata
KW - In vitro cell imaging
KW - Phycobiliprotein
KW - Physico-chemical properties
KW - R-phycoerythrin
KW - SDS-PAGE
UR - http://www.scopus.com/inward/record.url?scp=85121232561&partnerID=8YFLogxK
U2 - 10.1016/j.ijbiomac.2021.11.099
DO - 10.1016/j.ijbiomac.2021.11.099
M3 - Article
C2 - 34813785
AN - SCOPUS:85121232561
SN - 0141-8130
VL - 194
SP - 563
EP - 570
JO - International Journal of Biological Macromolecules
JF - International Journal of Biological Macromolecules
ER -