US HUPO 2025

It was great to be back in the US for the first time since ASMS 2024. This was also the first conference in which you got to meet Jenna, our Customer Experience Manager, and Zac and Erik, our Business Development Manager for North America and Head of Global Sales.

Danni delivered an excellent showcase of the performance gains Aurora Series columns can provide with some incredible data supplied by Karl Mechtler and Johannes Bruno Müller-Reif utilising the Aurora Rapid 8×75 and Aurora Rapid 8×150 respectively.

Monday, February 24 from 1:40 pm – 3:00 pm
Presented by Dr. Daniela-Lee Smith, Research & Development Scientist, IonOpticks

ABSTRACT

Proteomic analyses of large cohorts of human patient samples plays a critical role in discovering novel biomarkers for diagnosis of disease and responses to treatment. Here, we introduce a novel packed emitter chromatography column, the IonOpticks Aurora Rapid 8×150 (8 cm x 150 μm inner diameter, 1.7 μm C18), designed to improve the performance of large cohort proteomic analyses. Quality control samples embedded within a large human patient cohort were used to assess the efficiency and reproducibility of the column.

A HeLa tryptic digest (200 ng) was separated on Aurora Rapid 8 cm x 150 μm column on a 60SPD method using Evosep One and Thermo Fisher Orbitrap Astral Mass Spectrometer. MS parameters: MS1 m/z range(Orbi)/MS2(Astral) from 380 to 980, isolation windows of 2Th, a maximum injection time (IT) of 3 ms, FAIMS compensation voltage (CV) set at -40, resolution for MS1 set at 120,000. Patient plasma samples were analysed using 100 samples per day method with approximately 700 samples run between quality control (QC) injections. QC injections were included to monitor system performance (acquired using 60 sampler per day method).The data were processed using DIA-NN software (version 1.8.1) with match between runs enabled. 

The IonOpticks Aurora Rapid 8×150, paired to the EvoSep One and Thermo Fisher Orbitrap Astral Mass spectrometer achieved robust protein identifications, with over 9,300 proteins and 100,000 unique peptides identified per QC run. Coefficient of variation, Pearson correlation and average peak width were calculated to assess column reproducibility across the cohort. A median CV of 2.52 %, >0.98 Pearson correlation score and 2.9 second FWHM were achieved across the 257 QC analyses.

Our data demonstrates that the novel packed emitter column combined with the Evosep One and Thermo Fisher Orbitrap Astral Mass Spectrometer enabled maximum identifications whilst ensuring robust and reproducible protein quantification across a large patient plasma cohort analysis.

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