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. 2022 Sep 20;13:988870. doi: 10.3389/fpls.2022.988870

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Copyright © 2022 Kiefer, Niemeyer, Probst, Erkel and Schroda.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Enrichment of secreted SARS-CoV-2 spike protein from Chlamydomonas culture medium. (A) Attempted purification of octa-histidine-tagged PF/Fu-S protein from 250 mL culture medium via nickel affinity chromatography. Proteins in 1.7 mL of input (In), flow-through (FT), and washes (W1 and W2), and those in the entire eluate (E1-3) were precipitated with TCA. TCA precipitates and 20 μL of three elution fractions (E) were analyzed by immunoblotting. (B) Spike protein enrichment. Proteins in 100 mL of culture medium of wild-type (WT) and a PF/Fu-S producing transformant were precipitated with TCA (In, 85x enriched), concentrated by a rotary evaporator (In, 17x enriched) and centrifugal filters (34x enriched), or precipitated with (NH₄)₂SO₄ (In, 34x enriched). Enriched proteins were centrifuged to separate aggregated (P) and soluble (S) proteins. All proteins were analyzed by immunoblotting.