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PMC:7795856 / 25788-28012 JSONTXT

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LitCovid-PubTator

Id Subject Object Predicate Lexical cue tao:has_database_id
465 860-863 Gene denotes Tα1 Gene:134864
466 875-878 Gene denotes Tα1 Gene:134864
467 1050-1053 Gene denotes Tα1 Gene:134864
468 1304-1307 Gene denotes Tα1 Gene:134864
469 1573-1576 Gene denotes Tα1 Gene:134864
470 1683-1686 Gene denotes Tα1 Gene:134864
471 117-128 Chemical denotes citric acid MESH:D019343
472 392-408 Chemical denotes ammonium sulfate MESH:D000645
473 547-557 Chemical denotes AEX buffer
474 559-563 Chemical denotes sAEX
475 571-579 Chemical denotes Tris/HCl
476 586-590 Chemical denotes EDTA MESH:D004492
477 668-672 Chemical denotes sAEX
478 995-1006 Chemical denotes sAEX buffer
479 1120-1130 Chemical denotes CEX buffer
480 1138-1148 Chemical denotes Na-citrate
481 1217-1234 Chemical denotes TOYOPEARL Sulfate
482 1288-1291 Chemical denotes CEX
483 1383-1387 Chemical denotes NaCl MESH:D012965
484 1639-1643 Chemical denotes NaCl MESH:D012965
485 1749-1759 Chemical denotes AEX buffer
486 1767-1779 Chemical denotes ethanolamine MESH:D019856
487 1780-1783 Chemical denotes HCl MESH:D006851
488 1928-1938 Chemical denotes AEX buffer
489 2001-2005 Chemical denotes NaCl MESH:D012965
490 2105-2113 Chemical denotes peptides MESH:D010455
491 2165-2170 Chemical denotes water MESH:D014867
492 138-151 Disease denotes EmulsiFlex C5 MESH:C537005
493 1818-1840 Disease denotes mL Fractogel® EMD TMAE MESH:C562593

LitCovid-sentences

Id Subject Object Predicate Lexical cue
T148 0-4 Sentence denotes 4.3.
T149 5-34 Sentence denotes Purification of PASylated Tα1
T150 35-198 Sentence denotes Bacteria were harvested by centrifugation and disrupted in the presence of 100 mM citric acid using an EmulsiFlex C5 cell homogenizer (Avestin, Mannheim, Germany).
T151 199-354 Sentence denotes After centrifugation for 20 min (39,200× g) at 16 °C, the supernatant was filter-sterilized using a 0.45 µm syringe filter (Sartorius, Göttingen, Germany).
T152 355-499 Sentence denotes The clear filtrate was adjusted with ammonium sulfate to 30% saturation, stirred for 30 min at RT, and centrifuged for 30 min (39,200× g) at RT.
T153 500-688 Sentence denotes The precipitate was solubilized in subtractive AEX buffer (sAEX; 20 mM Tris/HCl, 1 mM EDTA, pH 8.5), filter-sterilized, and dialyzed twice against a 100-fold volume of sAEX buffer at 4 °C.
T154 689-838 Sentence denotes Subsequent chromatography steps were performed on an ÄKTA Explorer 100 system (GE Healthcare, Freiburg, Germany) operated at a flow rate of 5 mL/min.
T155 839-1007 Sentence denotes The dialyzed sample (Tα1-PAS or PAS-Tα1) was first applied to a 5 mL TOYOPEARL NH2-750F column (Tosoh Bioscience, Griesheim, Germany) equilibrated with the sAEX buffer.
T156 1008-1166 Sentence denotes The flow-through containing the PASylated Tα1 was collected and dialyzed twice against a 100-fold volume of the CEX buffer (20 mM Na-citrate, pH 3.0) at 4 °C.
T157 1167-1299 Sentence denotes The protein solution was then applied to an 85 mL TOYOPEARL Sulfate-650F column (Tosoh Bioscience) equilibrated with the CEX buffer.
T158 1300-1444 Sentence denotes PAS-Tα1 quantitatively bound to the column and the pure protein was eluted using a NaCl concentration gradient (0–250 mM over 2 column volumes).
T159 1445-1667 Sentence denotes In the case of Tα1-PAS, the CEX flow-through fraction contained the N-terminally acetylated protein, whereas the non-acetylated Tα1-PAS stayed bound to the column and could again be eluted in a NaCl concentration gradient.
T160 1668-1939 Sentence denotes The acetylated Tα1-PAS fraction from the flow-through was dialyzed twice against AEX buffer (20 mM ethanolamine/HCl, pH 10) and then applied to a 100 mL Fractogel® EMD TMAE (M)strong anion exchanger (Merck Millipore, Burlington, MA, USA) equilibrated with the AEX buffer.
T161 1940-2080 Sentence denotes This time, highly pure acetylated Tα1-PAS was eluted using a NaCl concentration gradient (0–200 mM over 2 column volumes) in the AEX buffer.
T162 2081-2224 Sentence denotes Both purified PASylated peptides were dialyzed 5 times against a 200-fold volume of water, lyophilized, and stored at −20 °C until further use.