2016/01/12 by Camille Lombard‐Banek, Sally A. Moody, Péter Nemes · 4 citations
Biochemistry, Genetics and Molecular Biology · Chemistry · #Advanced Proteomics Techniques and Applications #Biochemistry #Biology #Blastomere #Cell #Cell biology #Chemistry #Chromatography #Computational biology #Electrospray ionization #Embryo #Embryogenesis #Gene #Mass Spectrometry Techniques and Applications #Mass spectrometry #Molecular biology #Population #Proteome #Proteomics #Quantitative proteomics #Single-cell and spatial transcriptomics #Xenopus
paper · pdf · doi:10.1002/anie.201510411
openalex publication_date 2016/01/12 · openalex created_date 2016/06/24 · openalex updated_date 2026/08/06
We advance mass spectrometry from a cell population-averaging tool to one capable of quantifying the expression of diverse proteins in single embryonic cells. Our instrument combines capillary electrophoresis (CE), electrospray ionization, and a tribrid ultrahigh-resolution mass spectrometer (HRMS) to enable untargeted (discovery) proteomics with ca. 25 amol lower limit of detection. CE-μESI-HRMS enabled the identification of 500-800 nonredundant protein groups by measuring 20 ng, or <0.2% of the total protein content in single blastomeres that were isolated from the 16-cell frog (Xenopus laevis) embryo, amounting to a total of 1709 protein groups identified between n=3 biological replicates. By quantifying ≈150 nonredundant protein groups between all blastomeres and replicate measurements, we found significant translational cell heterogeneity along multiple axes of the embryo at this very early stage of development when the transcriptional program of the embryo has yet to begin.