PepSep Columns
PepSep Columns
HPLC & LC-MS CONSUMABLES

PepSep® Columns & Emitters

Everything you need for PASEF® speed
PEPSEP® COLUMNS & EMITTERS

HPLC Columns for Mega-sensitivity, High Throughput, and Maximum Reproducibility

High-performance liquid chromatography (HPLC) reversed phase columns are crucial components that determine the degree of separation in every analysis. In the world of proteomics, unlocking separation with precision can help with the quality of your analysis. 


Scientists, researchers, and clinicians desire robustness, speed, and sensitivity from their HPLC and mass spectrometry (MS) systems. Having a system that can accommodate large sample cohorts with minimal maintenance during chromatographic separation is a desirable trait to ensure sample reproducibility. The key to optimizing the chromatography process starts with columns. Having the right chromatographic column will assist in accurate data collection, reproducible results while offering higher throughput, and sensitivity.

 

This is where columns and tips are critical:
   (a) Bridging the gap between liquid chromatography (LC) and MS
   (b) Retaining peptides/ binding to active sites
   (c) Chromatographic resolution
    

omg OMx Podcast

omg OMx Podcast Episode 5 with Balyn Zaro

Tune in omg OMx Podcast where Dr. Balyn Zaro talks about PepSep® and more.

Flow with PepSep®

With a diverse portfolio of C18 nanoflow columns available in various lengths, inner diameters, and particle sizes, PepSep® ensures the perfect fit for every proteomics application.

PepSep® columns are crafted from industry-leading materials and are equipped with connectors to accommodate all chromatographic needs. Easily connect the columns to nanospray sources with a simple twist on and twist off design.

Spend less time swapping columns and more time on the important things, results. Amplify your experience when you combine PepSep® consumables with timsTOF series instruments, powered by Bruker ProteoScape™ to enhance your ability to acquire data at PASEF® speeds. 

"We are truly amazed by the performance of PepSep® columns, which have become indispensable for us in achieving remarkable high-throughput proteomics.

Working seamlessly alongside our Bruker timsTOF Pro mass spectrometer, PepSep® columns enable us to utilize higher nanoflow rates with minimal back pressure elevation. This translates to quicker analysis times and a substantial boost in sample throughput. "

David Gómez-Varela, Ph.D., Division of Pharmacology and Toxicology, University of Vienna, Vienna, Austria

PepSep® Advanced: Designed for Consistent, High-Quality Protein and Peptide Separation

Advanced columns delivering superior sensitivity and resolution for cutting-edge proteomics.

Key Features

  • Leading Sensitivity: Detects low-abundance peptides with maximum confidence
  • Enhanced Peak Sharpness: Achieves precise separations for improved data clarity
  • Comprehensive Identifications: Expands protein and peptide discovery capabilities
  • Uncompromised Robustness: Optimized flow dynamics and lower backpressure for smoother and faster operation

Technical Specification

  • Part No.: 1919826
  • Dimensions: 25 cm x 75 µm x 1.5 µm
  • Stationary Phase: C18
  • Column type: reversed phase
  • Pore size: 100 Å (1.5 µm)
  • Surface area: 400 m²/g (1.5 µm)
  • Temperature limits: 60°C
  • Ultra-High-Pressure inlet (up to 1000 bar)
  • Patented nanoConnect® outlet 
  • Flow rate: 100 - 500 nL/min
  • Applications: Single Cell Proteomics, Immunopeptidomics, General Proteomics, Plasma Proteomics, Analysis of PTM peptides, e.g., phosphopeptides

Developed for Ultimate Sensitivity and Enhanced Peptide Coverage

Identified unique peptide and protein groups, from 0.25, 1, 4 and 16 ng HeLa Tryptic Digest injection on an PepSep® Advanced column (25 cm x 75 µm x 1.5 µm) and Competitor A Column (pulled tip column, 25 cm x 75 µm x 1.7 µm). Samples were run on a nanoElute® 2 coupled to a Bruker timsTOF Ultra 2,  dia-PASEF® acquisition. Data analysed using DIA-NN 1.8.1.

> 5000 Protein Groups with 250 pg sample amount

Identified unique protein groups, from various amounts of K562 injections on an PepSep® Advanced  column (25 cm x 75 µm x 1.5 µm) using a 22 min active gradient. Samples were run on a nanoElute® 2 coupled to a Bruker timsUltra AIP, dia-PASEF® acquisition. Data analysed using DIA-NN 1.8.1 and K562 lib-based.

Achieves precise separations for improved data clarity

Retention times and peak widths of Biognosys iRT Kit peptides (1 µL injection volume, 1/300 dilution of stock solution) on a PepSep® Advanced column (25 cm x 75 µm x 1.5 µm) coupled to nanoElute® 2 and timsTOF Ultra, using a 22-minute active gradient.

"The new PepSep column has proven to be robust and consistently delivers reproducible chromatography. It produces sharper, more symmetrical peaks with no observable tailing, resulting in a 7–8% increase in identifications. Combined with the convenience of interchangeable emitters and competitive pricing, it has become our go-to column. It’s essentially plug-and-play."

Lynn Spruce, Technical Director, CHOP-Penn Proteomic Core Facility, Philadelphia, USA

PepSep® Classics: Versatile Solutions for Standard Proteomics Workflows

Reliable and versatile columns designed for routine proteomic analyses.

Key Features

  • Robust Durability: Engineered for consistent performance over extended use
  • Accelerated Throughput: Optimized for rapid analyses to increase laboratory efficiency
  • Exceptional Flexibility: Accommodates a wide range of separation tasks with ease
  • Cost-Effective Solution: Attractive pricing and longer lifespan with separated emitters 

Technical Specifications

  • Part No.: various, see table
  • Dimensions: various, see table
  • Stationary Phase: C18
  • Column type: reversed phase
  • Pore size: 100 Å (1.5 µm) or 120 Å (1.9 µm)
  • Surface area: 400 m²/g (1.5 µm)  or 300 m²/g (1.9 µm)
  • Temperature limits: 60°C
  • Ultra-High-Pressure inlet (up to 1000 bar)
  • Patented nanoConnect® outlet 
  • Flow rate: various, see table
  • Applications: various, see table

Choose from a wide range of columns feauturing various lenghts, inner diameters, and particle sizes:

Benefit Application Part No. Description Flow rate [nL/min]
Sensitivity Single Cell Proteomics, Immunopeptidomics, General Proteomics, High Sensitivity 1893627 25 cm x 50 µm x 1.5 µm 50 - 200
Sensitivity Single Cell Proteomics, Immunopeptidomics, General Proteomics, High Sensitivity 1893625 15 cm x 75 µm x 1.5 µm 150 - 750
Sensitivity Single Cell Proteomics, Immunopeptidomics, General Proteomics, High Sensitivity 1893626 15 cm x 50 µm x 1.5 µm 50 - 300
Sensitivity Single Cell Proteomics, Immunopeptidomics, General Proteomics, High Sensitivity 1895803 10 cm x 75 µm x 1.5 µm 150 - 1000
Sensitivity Single Cell Proteomics, Immunopeptidomics, General Proteomics 1893477 25 cm x 75 µm x 1.9 µm 100 - 600
Sensitivity Single Cell Proteomics, Immunopeptidomics, General Proteomics 1893473 15 cm x 75 µm x 1.9 µm 200 - 1000
Sensitivity Single Cell Proteomics, Immunopeptidomics, General Proteomics 1893472 10 cm x 75 µm x 1.9 µm 150 - 1500
Optimization General Proteomics, Plasma Proteomics, Mixed Samples, Gradient Versatility 1895838 40 cm x 150 µm x 1.5 µm 500 - 1000
Optimization General Proteomics, Plasma Proteomics, Mixed Samples, Gradient Versatility 1893479 25 cm x 150 µm x 1.9 µm 500 - 2000
Optimization General Proteomics, Plasma Proteomics, Mixed Samples, Gradient Versatility 1893471 15 cm x 150 µm x 1.9 µm 500 - 2000
Optimization General Proteomics, Plasma Proteomics, Mixed Samples, Gradient Versatility 1893474 15 cm x 150 µm x 1.5 µm 500 - 1500
Throughput General Proteomics, Plasma Proteomics, Mixed Samples, Gradient Versatility, High-Throughput 1893470 8 cm x 150 µm x 1.5 µm 500 - 5000
Throughput General Proteomics, Plasma Proteomics, Mixed Samples, Gradient Versatility, High-Throughput 1895619 4 cm x 150 µm x 1.9 µm 500 - 5000

PepSep® Classic 50 cm x 75 μm x 1.5 μm

The PepSep® Classic 50 cm x 75 μm x 1.5 μm is designed for long gradients, which are essential for spectral library generation. Spectral libraries play a crucial role in proteomics, particularly in mass spectrometry-based proteomics.

Different K562 concentrations were injected onto PepSep® Classic 50 cm x 75 μm x 1.5 μm column attached to the timsTOF HT coupled to the nanoElute 2 UHPLC. A standard DDA PASEF method, 1/k0 start 0.70, 1/k0 end 1.3 was used. results were searched using Bruker ProteoScape™ search engine.

PepSep® Classic 5 cm x 150 μm x 1.5 μm:
A Short Column for High-Throughput Proteomics

Designed for high-throughput proteomics, the PepSep® Classic 5 cm x 150 µm x 1.5 µm allows for gradients less than 15 minutes. This makes it ideal for applications where speed is essential, such as the analysis of clinical samples or the screening of drug libraries.

Results of different K562 concentrations separated on the PepSep®  Classic 5 cm x 150 µm x 1.5 µm column. The column was coupled to the nanoElute® and timsTOF HT. Two short gradients, 5 minutes and 10 minutes were used. The dia-PASEF 6x3 method was used with the IM set at 0.6 1.k0 start and 1.4 1/k0 end. Results were searched with DIA-NN.

PepSep® Classic 25 cm x 75 μm x 1.5 μm:
Enhance Sensitivity

Choosing the right column is difficult. A make-or-break decision that may increase protein and peptide identifications. The PepSep® Classic 25 cm x 75 µm x 1.5 µm C18 HPLC columns enable the analysis of a higher number of proteins and peptides from routine to restricted sample types. 

Based on the results from different digest concentrations of K562 run on the PepSep® Classic 25 cm x 75 µm x 1.5 µm over a 60 minute gradient. The PepSep® column was coupled with nanoElute® and timsTOF HT using a 6x3 dia-PASEF® window scheme and Bruker ProteoScape® to search the data in real-time.

PepSep® Classic 25 cm x 150 μm x 1.5 μm:
Greater Depth, Breadth and Rapid Proteomics

Core labs and academic labs want a single column solution for all applications to achieve deeper coverage, higher throughput, and better identification. 

PepSep® Classic 25 cm x 150 µm x 1.5 µm will help increase instrument performance, allowing scientists the liberty to have both sample throughput at PASEF® speeds and high 4D-sensitivity to dive deeper into the unknown and open the potential of the plasma proteome. 

Results from PepSep® Classic 25 cm x 150 µm x 1.5 µm columns on 45- and 60-min gradients with different K562 digest concentrations. The column was coupled with nanoElute® and timsTOF HT using a 6x3 dia-PASEF® window scheme and Bruker ProteoScape™ to search the data in real-time.

PepSep® Classic 25 cm x 150 μm x 1.5 μm:
Excellent Performance Driven by Enhanced 4D-Sensitivity

Rather than deciding between a shorter gradient and high throughput, PepSep® Classic 25 cm x 150 μm x 1.5 μm C18 HPLC columns offer you the best of both worlds. It’s time to improve your proteomics analysis with better coverage, throughput, sensitivity, and reproducibility. Pair the PepSep® Classic 25 cm x 150 μm x 1.5 μm with the timsTOF Pro 2 to find unprecedented proteomics depths — more than 5500 protein groups, 42,000 peptides in a 30 min gradient.

Using the nanoElute® UHPLC coupled to the timsTOF Pro, 200 ng K562 was run on the PepSep® Classic 25 cm x 150 μm x 1.5 μm with the 5 mm Thermo Pepmap trap. In this experiment, the default short gradient DDA MS method was used. The data was analyzed using MSFragger.

PepSep® Classic 10 cm x 150 μm x 1.5 μm:
Advancing Proteomics Research 

Scientists spend time fine tuning their methods to achieve broader coverage and higher throughput when mining plasma proteomes. 

The PepSep® Classic 10 cm x 150 µm x 1.5 µm increases instrument performance to help drive your analysis. When combined with the timsTOF HT, immerse yourself in 4D-Proteomics™, so you can accurately analyze 1000s of samples with high throughput at PASEF® speed.

PepSep® Classic 10 cm x 150 μm x 1.5 μm 20 min and 6 min: Precursors, peptides and protein groups identifications measured from healthy human donor plasma on the timsTOF HT. Plasma was prepared using the PreOmics iST-BCT kit. Peptide separation was performed on the nanoElute UHPLC coupled to the PepSep® MAX column run over 5.8- and 20-min gradients. 200 ng were injected for both gradients. Spectronaut 18 with directDIA+ was used for data analysis. 
DDA and DIA results of 200 ng commercial HeLa digest run on 50 samples per day (17.8 min gradient) using the PepSep® Classic 10 cm x 150 μm x 1.5 μm column attached to timsTOF HT.  For dia-PASEF®, a 6x3 windows scheme was used. Data results were generated in Bruker ProteoScape™ search engine.

 
Analyze More Samples, in Less Time, with Better Reproducibility

The proteomics landscape requires high throughput to capture meaningful rich data. PepSep® Classic 10 cm x 150 μm x 1.5 μm columns help you get there. It's time to improve your HPLCs capabilities. 

Identify, characterize and quantify peptides and proteins. Get excellent reproducibility, while outlasting leading columns up to 2x longer.

"At first, we were just using PepSep columns on our Bruker timsTOF Pro mass spectrometer. But we have been so impressed with their performance and affordable price point that we now use them across all instruments in our lab.  

 

These columns really hit a sweet spot of top-notch chromatography but with minimal back pressure. When compared to traditional nanospray columns, this allows us to use higher flow rates and minimize LC overhead time. This has been crucial for throughput, particularly when thousands of samples using shorter gradients."  

Danielle Swaney, Assistant Professor, Cellular Molecular Pharmacology, University of California, San Francisco, USA

When Performance Meets Simplicity 

Plug and play ready-to-use emitters are equipped with column connection union and spray tip to facilitate a hassle-free CaptiveSpray setup.

With two IDs available, PepSep® emitters cover the unique demands of proteomics applications. Compatible with all CaptiveSpray Sources.

Related Information

 

For Research Use Only. Not for use in clinical diagnostic procedures.