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. 2020 Dec 11;10:21779. doi: 10.1038/s41598-020-78711-6

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Analysis by SDS-PAGE and Reverse Phase (RP)-HPLC of RBD produced in P. pastoris or HEK-293T, and purified by NTA-Ni²⁺. Analysis of recombinant RBD fractions eluted from a NTA-Ni²⁺ column by 300 mM imidazole after purification from supernatants of a P. pastoris culture (A) or of HEK-293T cells (B). (C) RP-HPLC Analysis of RBD. Profiles for RBD produced in P. pastoris (red) and HEK-293T mammalian cells (black). The inset shows the expanded region of the chromatogram where the highest peaks eluted. The dashed blue line indicates the variation of acetonitrile (% v/v) during the experiments. Peaks 1, 2 and 3 from RBD produced in P. pastoris correspond to areas of 10.1, 2.8 and 87.1%, respectively.