Česká společnost pro hmotnostní spektrometrii

(13. ročník České konference hmotnostní spektrometrie a 11. Neformální proteomické setkání - ThO-08)
Low-input proteomics using the Orbitrap Astral

Pavel Talacko ^1 *, Karel Harant ^1,2, Jana Jašprová ¹

1. OMICS Mass Spectrometry Core Facility, Biocev, Faculty of Science, Charles University
2. Mass Spectrometry of biopolymers, IOCB Prague

Abstrakt

Proteomics has undergone a significant advancement in recent years, both in terms of sensitivity and the number of identified proteins. This progress has been driven by the widespread implementation of data-independent acquisition (DIA) and the emergence of new generations of mass spectrometers. The Orbitrap Astral represents a recent innovation within the Orbitrap platform, featuring a novel Astral analyzer characterized by exceptionally high sensitivity, rapid acquisition speed, and high resolving power. The introduction of this technology has markedly expanded the limits of detection in proteomic analyses, enabling the characterization of low-abundance protein populations that frequently play crucial roles in diverse cellular processes. Consequently, it substantially reduces the amount of sample material required for comprehensive analysis, which is particularly advantageous in single-cell proteomics.

At our facility, we have optimized a workflow for the analysis of sub-nanogram sample quantities, based on nano-flow HPLC coupled to the Orbitrap Astral mass spectrometer. Under these conditions, we are able to identify over 4,300 proteins from as little as 125 pg of cellular lysate, making single-cell proteomics experiments accessible to our users.

* Korespondující autor: pavel.talacko@natur.cuni.cz