Purification and desalting of peptides in bottom-up proteomic approaches.

About

BioSPE™ 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.

Benefits

• No storage constraint (store dry at room temperature)
• Different formats available to adapt to sample volume and throughput (StageTips, Spin columns, 96 wellplates and SPE cartridges)
• Two capacities available for each format depending on your peptide amount

Formats & Capacities

Details regarding the loading and elution volumes for our different BioSPE™PurePep 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™PurePep 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).

Video

Improvement of protein identification by LC-MS/MS in complex biological samples such as plasma by peptide fractionation at basic pH.

About

BioSPE™ PepFrac is used for peptide fractionation by hydrophobicity at basic pH. This fractionation method being orthogonal to peptide separation by reversed-phase liquid chromatography, sample complexity is reduced for deep proteome sequencing and quantitative analysis.

The BioSPE™ PepFrac kit is composed of:

• Fractionation columns based on an innovative reversed-phase resin
• Adapters (for tips centrifugation) and/or collection tools (centrifuge tubes or collection plates). Each kit contains nine collection tools (one for the conditioning/loading/washing steps and one for each elution fraction)
• A detailed protocol for fast and efficient peptide fractionation (up to 8 fractions)
• Triethylamine (TEA) solution

To determine which format and capacity will best suit your needs, please refer to the tables in the “Formats & Capacities” tab.

Benefits

• Increased number of identified proteins compared to unfractionated samples (up to 30% increase)
• Minimized fraction overlapping and good distribution of peptides in all fractions
• No desalting step required prior to LC-MS/MS analysis (contrary to fractionation by salt)
• No storage constraint (store dry at room temperature)
• Different formats available to adapt to sample volume and throughput (StageTips, Spin columns, 96 wellplates and SPE cartridges)
• Two capacities available for each format depending on your peptide amount

Formats and Capacities

Details regarding the loading and elution volumes for our different BioSPE™PepFrac 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™PepFrac products :

• For Standard capacity, product references contain the letter S (e.g. Tips-PepFrac.S8.200.96).
• For High capacity, product references contain the letter H (e.g. Tips-PepFrac.H8.200.96)

Removal of magnetic beads from samples after Single-Pot, Solid-Phase-enhanced Sample preparation (SP3).

About

BioSPE™ BeadRem is used to easily and quickly filter-out magnetic beads that remain in peptide or protein samples after SP3 procedure and that could plug the chromatographic column during LC-MS/MS analysis.

BioSPE™ BeadRem is available in two different versions, depending on the nature of your sample:

• BioSPE™ BeadRem Aqua for aqueous samples
• BioSPE™ BeadRem Orga for samples containing organic solvents

The BioSPE™ BeasRem kit is composed of:

• Filtration columns
• Adapters (for tips centrifugation) and/or collection tools (centrifuge tubes or collection plates). Each kit contains one collection tool to collect peptides/proteins in the flow-through
• A detailed protocol for fast and efficient removal of beads and recovery of peptides/proteins

To determine which format will best suit your needs, please refer to the table in the “Formats” tab.

Benefits

• Efficient removal of magnetic beads as small as 1µm diameter
• Fast and easy 3-step pass-through process
• Possibility to filter-out non magnetic beads such as agarose beads
• High peptide/protein recovery
• No storage constraint (store dry at room temperature)
• Different formats available to adapt to sample volume and throughput (StageTips, Spin columns, 96 wellplates and SPE cartridges)

Formats

Details regarding the loading volumes for our different BioSPE™ BeadRem products to find the best product depending on your sample volume.

Purification of intact proteins in top-down proteomic approaches.

About

BioSPE™ PureProt is used for the purification and desalting of intact proteins and large peptides (MW > 10kDa) from biological matrices (serum, plasma…) or food matrices (milk), prior to LC-MS/MS analysis.

The BioSPE™ PureProt kit is composed of:

• Purification 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 easy and efficient clean-up of proteins

To determine which format and capacity will best suit your needs, please refer to the tables in the “Formats & Capacities” tab.

Benefits

• High recovery yield
• No storage constraint (store dry at room temperature)
• Different formats available to adapt to sample volume and throughput (StageTips, Spin columns, 96 wellplates and SPE cartridges)
• Two capacities available for each format depending on your protein amount

Formats and Capacities

Details regarding the loading and elution volumes for BioSPE™PureProt 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™PureProt products :

• For Standard capacity, product references contain the letter S (e.g. Tips-PureProt.S.200.96).
• For High capacity, product references contain the letter H (e.g. Tips-PureProt.H.200.96)

Purification and enrichment of glycans and glycopeptides.

About

BioSPE™ GlycaClean can be used for the purification and enrichment of both glycopeptides and glycans, depending on your application.

The BioSPE™ GlycaClean kit is composed of:

• Enrichement 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 simple enrichment of glycopeptides or glycans

To determine which format and capacity will best suit your needs, please refer to the tables in the “Formats & Capacities” tab.

Benefits

• High recovery yield
• No storage constraint (store dry at room temperature)
• Different formats available to adapt to sample volume and throughput (StageTips, Spin columns, 96 wellplates and SPE cartridges)
• Two capacities available for each format depending on your protein amount

Formats and Capacities

Details regarding the loading and elution volumes for our different BioSPE™GlycaClean 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™GlycaClean products :

• For Standard capacity, product references contain the letter S (e.g. Tips-GlycaClean.S.200.96)
• For High capacity, product references contain the letter H (e.g. Tips-GlycaClean.H.200.96)

Efficient extraction and purification of metabolites and small molecules.

About

BioSPE™ PureOmics is used to desalt and concentrate small molecules and metabolites from different matrices (urine, plasma, culture medium…).

The BioSPE™ PureOmics kit is composed of:

• Purification 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 efficient desalting and extraction of metabolites and small molecules

Benefits

• High recovery yield on a broad polarity range
• No storage constraint (store dry at room temperature)
• Different formats available to adapt to sample volume and throughput (StageTips, Spin columns, 96 wellplates and SPE cartridges)

How to Choose the Reference

The following table will help you select the BioSPE™ PureOmics formats and references that are the most adapted to your metabolomic applications, depending on your sample volume.

Products list

Easy, fast and automatable filtration for protein precipitation.

About

AttractFiltra® Protein Precipitation plates make easy, fast and automatable, the removal of plasma/serum proteins prior to the analysis of small molecules present in these complex matrices.

This very important step is obtained by precipitating proteins in presence of an organic solvent (e. g. acetonitrile). In the 2mL format of each well, you can leave your organic solvent in the well for several hours without any leakage.

The addition of the serum/plasma and the subsequent mixing lead to the protein precipitation. Then, using a positive pressure or vacuum manifold or centrifugation, the filtrate containing small molecules is collected for a LC-MS/MS analysis. Usually the proportion is 1 volume of biological fluids to 3 volumes of organic solvent.

The overall process is very short and can be lead easily with an automate.

Products list

Isolation of circulating DNA et RNA on plasma and urine matrices.

About

BioSPE™ PureGenNucleo cfDNA is a ready-to-use kit for the efficient isolation, purification, and concentration of circulating cell-free DNA and RNA from human plasma or urine samples.

Circulating cell-free DNA (ccfDNA or cfDNA) is a fragmented DNA that freely circulates in the bloodstream, without being associated with cells. Though it is not always the case, this DNA can be tumor-derived. In this case, cfDNA mostly originates from apoptotic cells.

If cfDNA usual concentrations in plasma range from 0.1ng DNA per mL to several hundred ng per mL of plasma, the total amount of circulating DNA and RNA in urine or plasma depends on several criteria, including but not limited to:

• Health condition of the donor
• Sampling method
• Handling of the sample
• DNA isolation method…

Composition of the kit

BioSPE™ PureGenNucleo cfDNA mini spin columns kit is sold in two packaging sizes (10/pk or 50/pk) containing the following elements:

We also provide the following two accessories in the kit: Reservoirs (200mL) and manifold adapters.

Other formats available

BioSPE™ PureGenNucleo cfDNA kit is also available with 96 well plates, ideal for high throughput and automated processing.

Other formats will be available soon:

• SPE Tips for single cell analysis: ideal for low input material and small sample volume < 1mL, with simple processing by centrifugation
• 15mL spin columns: ideal for large sample volumes up to 20mL
  Don’t hesitate to contact us with any requests!

Specifications and Benefits

• High recovery of fragmented DNA (≥ 50bp) and RNA
• Convenient processing of up to 5mL of sample at once thanks to the adapted reservoirs
• Small final elution volume (60µL) for highly concentrated DNA
• Purified DNA and RNA compatible with downstream PCR or cancer biomarker analysis
• Processing: vacuum (+ centrifugation for elution)
• Use: for research use only

Products List

In order to prepare high quality peptide samples for LC-MS analysis, it is very important to ensure excellent overall quality of proteomic experiments. Peptide samples collected after enzymatic digestion require treatment to remove salts, possible pieces of gels or particles that can damage the equipment or falsify the results.

AttractSPE®Tips is the ideal tool for this application as well as for drug extraction or small molecule analysis in urine or serum in bioanalysis.

Several capacities are available to help you in your purification protocols.

Our application notes have shown that our SPE tips C18 are the perfect tools for peptides desalting with excellent results.

SPE tips or Stagetips, centrifugeable, compatible with automates for high throughput screening, useful for biomolecular applications of peptide desalting, proteomics, and biomarker discovery.

About and Capacity

About

Process of use: centrifugation, positive pressure.
Our SPE tips can’t be used with a pipette.

Formats: 10µL, 200µL and 1mL.

For any other format, contact us.

NEW: eco packaging of 960 tips in bulk now available for AttractSPE® Tips C18!

• More economical: the bulk packaging represents a saving of up to 30% compared to the price of 10 boxes of 96 tips!
• More ecological: new packaging is made with 90% less plastic and is 100% recyclable!

Contact us for more details!

Capacity

Each product is available with different capacities, which are designated T1, T2 or T3 in the product designations. This designation refers to the layer thickness of SPE tips. For a more reliable product, each tool contains only ONE layer.

Table legend: T1, T2, T3 = maximum loading capacity (µg)

Video and Webinar

Watch in replay our webinar on Phosphoproteomics with AttractSPE®Disks Tips!

1mL 96 wellplates for SPE for high throughput purification/desalting or fractionation by automate.

About

AttractSPE® Disk 96 Plate is a plate with 96 SPE columns containing immobilized SPE disks that enables a high throughput clean-up with the simultaneous preparation of 96 samples. Thanks to this small sorbent amount and a high efficiency, almost 100% of the original sample is recovered. The 1mL well contains a 10-20mg SPE disks for a high capacity. Due to the weak dead volume of the solid phase, a very small elution volume is required.

Benefits

• All sorbents available on catalog or on demand
• Several sorbent weights available
• Easy handling with automates or liquid handling robots – spinnable
• AttractSPE® Manifold for 96 wellplate format or AttractSPE®Positive pressure Manifold for 96 wellplate
• Capacity up to 500µg for T1, 1mg for T2 and 1.5mg for T3

Format and Capacity

Table legend: T1, T2, T3 = maximum loading capacity (µg)

Products List

Combined advantages of 96 well plates and SPE tips for high throughput clean-up on StageTips by automation:

• High-capacity SPE disks sorbent

• Highly efficient

About

AttractSPE® Disks 96 Plate for microelution is a plate with 96 microSPE columns containing immobilized SPE disks that enables a high throughput clean-up with the simultaneous preparation of 96 samples. Thanks to this small sorbent amount and a high efficiency, almost 100% of the original sample is recovered. Each well of 400µL contains 1-2mg of SPE disks with a pipette tip shape. This is the ideal product to carry out 96 stagetips experiments in one step, enabling fractionation and desalting with a very high speed.

Benefits

• All sorbents available on catalog or on demand C18, C8, C4, SAX, SCX, SDB, HLB, RPS
• Several sorbent weights available
• Compliant with centrifugation, positive pressure and vacuum manifolds
• Easy handling with automates or liquid handling robots – spinnable
• AttractSPE® Manifold for 96 well Plate format or AttractSPE® Positive pressure Manifold for 96 well Plate
• Capacity up to 35µg for T1, 70µg for T2 and 105µg for T3

Format and Capacity

Table legend: T1, T2, T3 = maximum loading capacity (µg)

Product List

96 SPE well plates with powder sorbents (polymer, silica-based or molecularly imprinted polymers) for high throughput analysis or screening.

About

Most products using a powder sorbent in the portfolio of SilactSPE™, AttractSPE® or AFFINIMIP®SPE can be available as 96 SPE well plates of 1mL or 2mL.

Benefits

• Most sorbents available on catalog or on demand (SilactSPE™, AttractSPE® or AFFINIMIP®SPE)
• Several sorbent weights available
• Compliant with automate used, positive pressure manifold or 96 wellplate vacuum manifold

1mL and 2 mL collection plate compliant with automates

Products list

A SPE spin column for a high efficiency purification/desalting/fractionation of peptides/proteins/DNA/RNA or small molecules.

About

Thanks to their strength, monolithic SPE disks do not require frits to form the column bed (unlike traditional SPE products using powder). This allows recovery yields close to 100% after a serie of short centrifugations.

AFFINISEP proposes a range of 4 sizes spin tubes to make possible the scale-up of your purification thanks to similar disks (2mL microspin, 2mL minispin, 15mL spin and 50mL spin tubes).

They are commonly used for peptides/proteins/DNA & RNA purification and desalting or small molecules extraction.

Benefits

• 4 sizes of SPE spin format: 2mL microspin, 2mL minispin, 15mL spin and 50mL spin tubes
• Fast and easy extraction by centrifugation
• High-flow purification and high capacity
• 3 different capacities up to several mg for an easy scale-up
• For peptides/proteins/DNA & RNA purification and desalting or small molecules extraction
• Diversity of sorbents: C18, C8, C4 wide pore, SCX, SAX, SDB, HLB, RPS

Formats and Capacitites

Table legend: T1, T2, T3 = maximal loading capacity (µg)

Solid Phase Extraction Disks for Waters and Environmental Liquids Analysis at High Flow Rates.

For all your environmental SPE needs, AFFINISEP designed and developed AttractSPE®Disks Environment.

About

Our Innovation puts at your disposal a broad variety of sorbents (C18, SDB, HLB, RPS, Anion and cation exchanges). SPE Disks manufactured by AFFINISEP enable the loading of great volumes of waters. The treatment of environmental samples implies great volume of environmental liquids, that’s AttractSPE®Disks are especially designed for any kind of waters, dirty or ground waters. Despite its thinness, AttractSPE®Disks Environment remain dense and resistant during your sample analysis. AttractSPE®Disks Environment cover the extraction of a large scope of contaminants such as pesticides, PAHs or drug residues and is compliant with EPA methods.

AttractSPE®Disks Environment is also the perfect membrane to use for the passive sampler SPE Disk based or Chemcatcher thanks to a very good hold and ease to use.

AttractSPE®Prefilter can be added on top of the AttractSPE®Disk to prevent clogging in case of samples with a lot of particles.

For the extraction of molecules, we propose one, three or six-station filtration manifolds in our product catalog.
To help you, we have also developed an interesting kit, containing a Manifold and a box of disks. For more information, go to our “Kits for method development” tab.

Video

Guide Selection

Kits for Method Development

AttractSPE®Disks are outstanding concentration and sampling tools. To implement these methods in your laboratory, a manifold is required.

For this development phase, we offer special prices for 1 development kit:

Dev-Disks-47-MAN-1 is a development kit containing one Manifold – 1 station for disks 47mm supplied with one box of 20 AttractSPE®Disks of your choice – 47mm.

AttractSPE® Disks C18, SAX, SCX, RPS… for bioanalytical and biomolecular applications, DNA&RNA, peptides and proteins purification, fractionation and desalting, drug analysis, metabolomics…

About

As protein analysis has become more complex and more sensitive, the need for adequate materials meets more specific requirements. Proteomic sample preparation should be a critical consideration.

AFFINISEP brings to you a full range of membranes with AttractSPE®Disks BioMol with C18, C8, C4, SCX, SAX, RPS….

These SPE membranes are especially designed and made by AFFINISEP for your bioanalytical needs and to help you to develop your own tools for your R&D projects, for instance to make Stagetips by punching the membranes. Our high quality SPE membranes allow high throughput (HTS) for protein desalting or peptides purification.

AttractSPE®Disks BioMol are fully compatible with automated liquid handlers. Indeed, with these Solid Phase Extraction Disks, you can design the applications you wish such as removing impurities from samples and isolation of analytes from biomatrices with small elution volumes. Fast and efficient purification of peptides through Solid Phase Extraction is easily accessible.

Benefits

• For peptides/proteins/DNA & RNA purification and desalting or small molecules extraction
• Diversity of sorbents C18, C8, C4, SCX, SAX, RPS
• Easy adaptation to Bioanalytical formats
• Handling of tiny and viscous samples
• High density of homogeneous particles
• Compatible with automated liquid handlers and high throughput analysis (HTS)
• High capacity
• Several biotools are available using these SPE disks (Stage-tips, Spin columns, 96 well plate for microelution, cartridges)

Articles

• The HUSH complex represses retroviruses, transposons and genes to maintain the integrity of vertebrate genomes. In this paper, it was demonstrated that TASOR, the central HUSH subunit, is a multifunctional pseudo-PARP that directs HUSH assembly and epigenetic regulation of repetitive genomic targets.Clean-up of tryptic digests of proteins extracted from TASOR cells was performed with AttractSPE® Disks Bio RPS (SPE-Disks-Bio-RPS-M-47.20) packed into 200 µL pipette tips (equivalent to our AttractSPE® Disks Tips RPS, ref Tips-RPS-M.T1.200.96) prior to mass spectrometry analysis.TASOR is a pseudo-PARP that directs HUSH complex assembly and epigenetic transposon control, Christopher H. Douse, Iva A. Tchasovnikarova, Richard T. Timms, Anna V. Protasio, Marta Seczynska, Daniil M. Prigozhin, Anna Albecka, Jane Wagstaff, James C. Williamson, Stefan M. V. Freund, Paul J. Lehner, Yorgo Modis, Nature Communications, 2020, https://doi.org/10.1038/s41467-020-18761-6.

• Clathrin-mediated endocytosis (CME) is a central pathway for the internalization of proteins from the cell surface, thereby contributing to the maintenance of the plasma membrane protein composition. In this article, it was hypothesized that blocking AP-2 function, a key component for the formation of endocytic clathrin-coated vesicles (CCVs), and thus inhibiting CME may be a novel approach to identify new druggable targets, or to increase their residence time at the plasma membrane, thereby increasing the probability for efficient therapeutic intervention. Stage tips packed with two layers of AttractSPE® Disks Bio RPS (SPE-Disks-Bio-RPS-M-47.20) and equivalent to our AttractSPE® Disks Tips RPS (Tips-RPS-M.T2.200.96) were used for peptides desalting after tryptic digestion.Inhibition of clathrin-mediated endocytosis by knockdown of AP-2 leads to alterations in the plasma membrane proteome, David Tobys, Lisa Maria Kowalski, Eva Cziudaj, Stefan Müller, Peter Zentis, Elke Pach, Paola Zigrino, Tobias Blaeske, Stefan Höning, Traffic, Volume 22, Issue 1-2, 2021, https://doi.org/10.1111/tra.12770.