Vacuum Insulated Probe Heated Electrospray Ionization Source Enhances Microflow Rate Chromatography Signals in the Bruker timsTOF Mass Spectrometer.

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Journal of proteome research


washington; isb; captivespray (CS); data-independent acquisition-parallel accumulation and serial fragmentation (dia-PASEF); electrospray ionization (ESI); microflow liquid chromatography (μLC); slice-parallel accumulation and serial fragmentation (Slice-PASEF); vacuum insulated probe heated electrospray ionization (VIP-HESI) source; Humans; Animals; Mice; Spectrometry, Mass, Electrospray Ionization; Reproducibility of Results; Proteomics; Vacuum; Chromatography, Liquid; Peptides; Ions; Proteome


By far the largest contribution to ion detectability in liquid chromatography-driven mass spectrometry-based proteomics is the efficient generation of peptide molecular ions by the electrospray source. To maximize the transfer of peptides from the liquid to gaseous phase and allow molecular ions to enter the mass spectrometer at microspray flow rates, an efficient electrospray process is required. Here we describe the superior performance of newly design vacuum insulated probe heated electrospray ionization (VIP-HESI) source coupled to a Bruker timsTOF PRO mass spectrometer operated in microspray mode. VIP-HESI significantly improves chromatography signals in comparison to electrospray ionization (ESI) and nanospray ionization using the captivespray (CS) source and provides increased protein detection with higher quantitative precision, enhancing reproducibility of sample injection amounts. Protein quantitation of human K562 lymphoblast samples displayed excellent chromatographic retention time reproducibility (


Institute for Systems Biology