nanoElute^® 2

Built on the core principles of its predecessor, the nanoElute^® 2 has been refined for unrivalled robustness, ultimate chromatographic performance, and ease-of-use. 

• Unprecedented pump reproducibility and robustness 
• Ultra-sharp peaks for the best proteomics performance 
• Flexible user-controlled features and injection modes
• User friendly interface for early-adopters 
• Assisted troubleshooting procedures for expert users 
• Automatable sample preparation workflows 
• Reach extreme LC-MS sensitivity for “label-free single-cell level” workflows

The nanoElute^® 2 covers a broad range of applications from single-cell to high-throughput routine proteomics. This next generation system provides you with the highest chroma­tographic resolution resulting in exceptional peptide identification and quantification.

Unprecedented reproducibility and robustness achieved with: 

• New, hardened switching valves, for extended lifetime 
• Improved pump design 
• Finger-tight, torque-limiting capillaries, for leak-free, zero dead volume connections, without risk of overtightening or damaging capillaries.

The nanoElute^® 2 provides precise and reproducible gradient separation for different gradient lengths, resulting in narrow peak widths and excellent peak capacity. 

In combination with the timsTOF HT reliable performance was achieved for all injected quantities (see Figure C). This within a 41 min runtime method for 1600 ng K562 tryptic digest by data dependent acquisition parallel accumulation serial fragmentation PASEF^® (DDA).

With its unique valve design the nanoElute^® 2 enables software-controlled flow paths for dynamic inclusion of the trap column and backflushing the sample for sharper peaks. It retains the top desirable functionalities of its predecessor ensuring: 

• Dynamic on-the-fly method configuration 
• Efficient, predefined column washing and re-equilibration 
• Fully integrated and intelligent software-control
• Built-in quality control and leak management 
• Seamless switch between trap-and-elute, direct load and direct infusion experiments 

These features reduce the need to compromise between high performance, flexibility and robustness, therefore giving you full confidence in your results!

Fully automated self-testing and diagnosis is embedded into a user-friendly interface for early adopters, as well as assisted troubleshooting procedures for experts. 

New procedures allow for step-by-step evaluation of flow-paths to readily locate potential issues for easy troubleshooting and maintenance. 

Sample preparation procedures provide flexibility for user-defined performance evaluation, autosampler washing, and solvent exchange procedures.

Automatable sample preparation procedures alleviate the need for manual intervention can be configured based on user-defined input settings. The following liquid handling functionalities provide the means to dissolve and mix samples just prior to LC-MS analysis:

• Add from vial
• Dissolve
• Derivatize

The timsTOF SCP in combination with the nanoElute^® 2 UHPLC system achieves unprecedented sensitivity. About 4000 protein groups and over 22,000 precursors were identified from 1 ng of HeLa using a 80 min runtime method (i.e. gradient length of 66 min) and a data-independent acquisition parallel accumulation serial fragmentation (dia-PASEF^®).

Use the nanoElute^® 2 and the timsTOF SCP for end-to-end single cell label-free proteomics workflows. The nanoElute^® 2 injects directly from the label-free proteoCHIP holder used in the cellenONE cell sorter. In this workflow up to 144 single cells can directly be transferred from the cellenONE to the nanoElute^® 2 autosampler, after sorting and preparation, without pipetting steps, for ultra-sensitive proteomics analysis using the timsTOF SCP and TIMS-DIA-NN.

Assessment of sample pick-up of 250 pg and 1 ng in 1 μL total volume, and 2 ng in 2 μL total volume of peptides from a HeLa cell lysate digest directly injected from wells of the label-free proteoCHIP showed excellent reproducibility in protein group and peptide identification rates. Injections from the label-free proteoCHIP and from an autosampler vial demonstrated comparable results on protein group and peptide identification rates at different peptide concentrations and volumes.