Peptide desalting
Purification and desalting of peptides in bottom-up proteomic approaches with BioSPE® PurePep.
BioSPE® for peptide desalting (PurePep) is used for efficient peptide desalting after enzymatic digestion, prior to LC-MS/MS analysis. Up to 97% proteins are identified after desalting of HeLa digest on BioSPE® PurePep, with a good recovery of peptides on the whole polarity range, and good repeatability (RSD < 3%).
For desalting of the most hydrophilic peptides, we recommend the use of BioSPE® PurePep Broad, that can efficiently retain small polar peptides. BioSPE® PurePep Broad is also stable under extreme pH conditions (pH 0-14) and can run dry during the SPE process without consequences on your results.
The BioSPE® PurePep and BioSPE® PurePep Broad kits are composed of:
- Desalting columns
- Adapters (for tips centrifugation) and/or collection tools (centrifuge tubes or collection plates). Each kit contains two collection tools (one for the conditioning/loading/washing steps and one for the elution step)
- A detailed protocol for fast and easy desalting
To determine which format and capacity will best suit your needs, please refer to the tables in the “Formats & Capacities” tab.
- No storage constraint (store dry at room temperature)
- Different formats available to adapt to sample volume and throughput (SPE Tips, Spin columns, 96 & 384 wellplates and cartridges)
- Two capacities available for each format depending on your peptide amount
Details regarding the loading and elution volumes for our different BioSPE® Peptide desalting products to find the best product depending on your sample volume.
Information table regarding the capacities (maximal amount of peptides/proteins that can be retained) of our different BioSPE® Peptide desalting products :
- For Standard capacity, product references contain the letter S (e.g. Tips-PurePep.S.200.96),
- For High capacity, product references contain the letter H (e.g. Tips-PurePep.H.200.96).
Literature
Hijazi, H. et al. (2025) ‘Mind Your Spectra: Points to be Aware of When Validating the Identification of Isobaric Histone Peptidoforms’, Journal of Proteome Research.
In their latest publication, Hassan M. Hijazi et al. investigate the identification of mouse-specific histone variants H3mm7 and H3mm13 from brain and testis extracts using LC-MS/MS.
To enable accurate detection of post-translational modifications such as acetylation and lactylation, peptide purification was performed with Affinisep BioSPE® PurePep, which proved highly effective in removing interfering substances and preserving modified peptides.
This approach improved MS/MS spectral quality, particularly for critical ions like b1 and CycIm, and helped address challenges related to isobaric variants caused by uncontrolled modifications such as propionylation.
Product used : BioSPE® PurePep Tips 200µl (Catalog # KT-Adapt-Tips-PurePep.H.200.96)
Application notes
- Proteomics
AN-0019-01 | Estimation of the working range on BioSPE® PurePep SPE Tips – Standard capacity
Peptide desalting with BioSPE™ PurePep
Peptide fractionation
Improvement of protein identification by LC-MS/MS in complex biological samples such as plasma by peptide fractionation at basic pH.
Magnetic beads removal
Removal of magnetic beads from samples after Single-Pot, Solid-Phase-enhanced Sample preparation (SP3).
Intact protein purification
Purification of intact proteins in top-down proteomic approaches.
Products