nanoElute^®

The system performance is maintained through 3 layers of protection:

1. Automatic initialization, where the autosampler and pumps automatically run a preparation protocol before sample injection ensuring performance from the first to the last sample.
2. Built-in scheduling of preventive maintenance jobs based on usage patterns and integrated system expertise where the software will suggest how to optimize continued system operation.
3. Precise system diagnostics that automatically detects leaks and blockages and highlights them in the graphical user interface.

Tailor-made instrument methods are automatically generated using the Instant Expertise™ method editor, based on experimental mode, column selection and analysis time, delivering:

• LC-MS/MS method with intelligent, self-optimizing LC separation for higher productivity and expert-caliber results the first time.
• Optimal experimental conditions are automatically calculated from column type and sample characteristics to suit all proteomics samples
• Chromatographic resolution is preserved for best simultaneous ID and Quant capabilities

Gradients can be readily modified but most other parameters are hidden although they can still be edited in “expert” mode.

The system is preconfigured with a standard layout and a novel valve solution to ensure ease-of-use and system stability. The unique valve design ensures that the user can execute three different types of experiments without hardware changes and software controlled flow paths exist for using a trap column, bypassing the trap column or doing direct infusion for fast analyses of simple samples.

The unique valve design and novel instrument configuration enable:

• Software controlled flow paths
• Dynamic inclusion of trap column
• Backflushing the sample for sharper peaks

New nano-columns and a unique back-flushable trap cartridge holder complement the instrument and are recognized by the Instant Expertise algorithm.

Compact and dedicated designwith market proven autosampler and large volume, single stroke piston pumps support longer gradients and a wide flow range for efficient Omics knowledge generation.

Modern nano-ESI mass spectrometry (MS) analysis is putting increased pressure on the front-end LC separation to match the MS in terms of robustness and reliability as well as demanding the absolute highest quality and reproducibility of sample separation.

To ensure good individual system performance as well as to deliver consistency over multiple systems and across different labs, deeper system integration and novel approaches to quality control as well as intelligent system diagnostics are called for. This in turns requires rigorous protocols and automation for all phases of the experiment, driving new approaches to user interface design and data monitoring, as well as a critical rethink of all flow paths and liquid handling parts of the configuration.

The Bruker nanoElute^®is designed with years of proteomics know-how and nano-UHPLC expertise.

Based on industry leading components coupled with innovative solutions to common problems, it delivers trouble-free operation from the moment of installation through years of Omics knowledge generation.

When paired with the ultra high-resolution / accurate mass QqTOF, the Bruker impact II and the market proven CaptiveSpray ionization source, the nanoElute^®system is unbeatable when it comes to:

• Data-dependent and data-independent bottom-up proteomics
• Proteoform profiling for biomarker discovery and validation
• Intact protein characterization
• Glyco- and phosphoproteomics

The ultra high-resolution, accurate mass QqTOF, the Bruker impact II and the market proven CaptiveSpray ionization source are the perfect companions for the nanoElute^®system.

The complexity of the nanoElute^®’s built-in intelligence and applied chromatographic understanding is hidden behind a user interface that is intuitive and easy to use, but also powerful and flexible to operate.

Trouble-free Omics knowledge generation
The nanoElute^®system provides excellent chromatographic performance even for very short gradients in both separation modes. With a trap column installed, the focus is on speed and robustness, however the chromatographic performance is maintained due to new nano-columns and unique back-flushable trap cartridge.

Without a trap column installed, ultimate separation power is delivered where the trap column is elegantly by-passed by virtue of the novel valve geometry.

The unique instrument configuration allows the user to analyze samples in both “with trap” and “without trap” mode in the same sample batch without physically changing the hardware setup.

Separation of complex samples with a long column and gradient has been demonstrated both with and without trap column. The retention time and area reproducibility is excellent displaying suitability for all proteomics applications, including the most demanding ones.

Method
Predigested Bovine Serum Albumin (Bruker, Billerica, MA) was used as a representative for a simple proteomics sample (25 fmol/µL).
HeLa cell lysate was prepared according to Beck et al.¹ and acidified with formic acid to a final concentration of 200 ng/µL. For the simple sample, the tryptic peptides were separated on a novel nano-UHPLC system using a 10 min reversed phase gradient (2% - 35% solvent B in 10 min - 0.1% FA in ACN), on standardized 15cm 75µm bore column with C18 resin. The HeLa sample was separated with a reversed phase gradient (from 2% -24% in 90 min, 24-48 in 10 min solvent B - 0.1% FA in ACN), on a standardized 40cm 75µm bore columns with C18 resin. Data was acquired with an impact II QTOF MS (Bruker Daltonik, Bremen, Germany) equipped with a CaptiveSpray ion source..

References
[1] Beck, S., Michalski, A., Raether, O, Lubeck, M., Kaspar, S., Goedecke, N., Baessmann, C., Hornburg, D, Meier, F., Paron, I., Kulak, N.A., Cox, J. and Mann, M. “The Impact II, a Very High-Resolution Quadrupole Time-of-Flight Instrument (QTOF) for Deep Shotgun Proteomics.” Mol Cell Proteomics. 2015 Jul; 14(7): 2014-29.

Direct infusion for fast analysis

The unique valve design also allows ”Direct Infusion” for fast runs of simple samples e.g. BSA at 2 μL/min.

Direct infusion for fast analysis
The unique valve design also allows ”Direct Infusion” for fast runs of simple samples e.g. BSA at 2 μL/min.