Newomics MnESI-MS platform has been optimized for native MS. It directly addressed the aforementioned challenges of static nanospray ESI-MS and high flow ESI-MS for native MS.

The Newomics MnESI-MS platform has been interfaced to Thermo Fisher Orbitrap MS, Bruker timsTOF, and Agilent Q-TOF (beta), mass spectrometers, resulting in a unique and complete solution for native MS.

Newomics solution benefits:

• Better ionization: more charge available for each smaller droplet (via multinozzle) during electrospray ionization process
• Lower voltage compared to high flow: softer ionization (to maintain the protein, protein-drug interactions, and protein complexes in the native states)
• Room temperature for desolvation gas: maintain the protein, protein-drug interactions, and protein complexes in the native states)

Application note: Microflow LC-MS Analysis of Native Monoclonal Antibodies

Newomics® M3 emitters have achieved high throughput, high sensitivity, and robustness in native MS studies of monoclonal antibodies (mAb) and protein complexes. The M3 emitter has shown significant improvement compared to the traditional high-flow SEC-MS platform using HESI:

1. More than 10-fold sensitivity
2. Better preservation of the native conformation of antibodies
3. The flexibility of upstream and downstream analysis of mAb including flow splitting for simultaneous detection of mAb by UV and MS

Figure 1. Sensitivity comparison of M3 emitters with other emitters at different flow rates. A. Comparison of microflow SEC-MS using M3 emitters at 5 μl/min and high-flow SEC-MS using HESI at 200 μl/min. Shown here are the extracted ion chromatograms (m/z 4,500-6,000) from 50 ng IgG (i) and 200 ng IgG (ii), respectively. B. Comparison of multinozzle M3 emitters to single-nozzle emitters at microflow. (i). sensitivity comparison of M3 emitters with NanoTip, PicoTip, and HESI using the same microflow SEC column. (ii). LOD analysis for M3 emitters using serial dilution of IgG. The LOD is below 10 ng.

Application note: Native Protein Complexes analyzed with MnESI-MS platform

The MnESI platform demonstrated a significant gain of approximately 50-fold in sensitivity compared to the conventional HESI probe analytical flow method. With the sensitivity improvement from the MnESI platform, it is possible to analyze samples with a smaller amount or shorter gradient. Furthermore, reduction of high salt and protein complexes injection into the mass spectrometer minimizes the contamination to the instrument, enhances method robustness, and reduces the downtime due to cleaning.

Figure 2. Native SEC-LC/MS analysis of GroEL tetradecamer protein complex. a. TIC of GroEL 14-mer (6 μM) from a 20 min SEC-LC/MS run. b. Native MS1 analysis of GroEL 14-mer using the MnESI platform at microflow of 5 μL/min. Secondary complex of 15-mer was clearly detected. c. Native MS1 analysis of GroEL 14-mer using the Ion Max source with HESI probe at analytical flow of 50 μL/min. d. Native MS1 analysis of GroEL 14-mer (1.25 μM) using static NanoESI-MS. e. Sensitivity comparison of MnESI platform vs. HESI probe method.

Application Note: Monoclonal Antibodies analysis under Native conditions

Monoclonal antibodies (mAbs), such as therapeutic antibodies, antibody-drug conjugates (ADCs), biosimilars, and bispecific mAbs, have all been routinely analyzed using native MS. High-flow size exclusion chromatography (SEC)-MS may not be sensitive enough to detect proteins with low quantities due to low ionization efficiency at the high (analytical) flow rate. In this Application Note, we demonstrate a new microflow LC-MS platform for native MS of mAb, using Newomics® M3 emitters interfaced with a Thermo Fisher Orbitrap Q Exactive Plus mass spectrometer.