Benchmarking of Two Peptide Clean-Up Protocols: SP2 and Ethyl Acetate Extraction for Sodium Dodecyl Sulfate or Polyethylene Glycol Removal from Plant Samples before LC-MS/MS

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Authors

MARTINKOVÁ Petra KONEČNÁ Hana GINTAR Petr KRYŠTOFOVÁ Karolína POTĚŠIL David TRTÍLEK Martin ZDRÁHAL Zbyněk

Year of publication 2023
Type Article in Periodical
Magazine / Source INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
MU Faculty or unit

Central European Institute of Technology

Citation
Web fulltext
Doi http://dx.doi.org/10.3390/ijms242417347
Keywords peptide clean-up; magnetic beads; SP2; ethyl acetate extraction; sodium dodecyl sulfate; detergent; polyethylene glycol; Arabidopsis thaliana; LC-MS/MS
Description The success of bottom-up proteomic analysis frequently depends on the efficient removal of contaminants from protein or peptide samples before LC-MS/MS. For a peptide clean-up workflow, single-pot solid-phase-enhanced peptide sample preparation on carboxylate-modified paramagnetic beads (termed SP2) was evaluated for sodium dodecyl sulfate or polyethylene glycol removal from Arabidopsis thaliana tryptic peptides. The robust and efficient 40-min SP2 protocol, tested for 10-ng, 250-ng, and 10-µg peptide samples, was proposed and benchmarked thoroughly against the ethyl acetate extraction protocol. The SP2 protocol on carboxylated magnetic beads proved to be the most robust approach, even for the simultaneous removal of massive sodium dodecyl sulfate (SDS) and polyethylene glycol (PEG) contaminations from AT peptide samples in respect of the LC-MS/MS data outperforming ethyl acetate extraction.
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