1. ABOUT THE DATASET ------------ Title: Data supporting 'Ultrahigh-throughput and low-volume analysis of intact proteins with LAP-MALDI MS' Creator(s): Bob Challen, Rainer Cramer Organisation(s): University of Reading Rights-holder(s): University of Reading Publication Year: 2023 Description: As part of the PhD project 'Novel applications of liquid MALDI MS for the analysis of large biomolecules' funded by the University of Reading and Waters Corporation, the use of high-throughput analysis towards intact mid-sized proteins was investigated. Raw and processed data for liquid atmospheric-pressure matrix-assisted laser desorption/ionisation (LAP-MALDI) mass spectrometry analysis of three mid-sized proteins at analysis rates of up to 20 samples/second are presented. Cite as: Challen, Cramer (2023): Ultrahigh-throughput and low-volume analysis of intact proteins with LAP-MALDI MS. University of Reading. Dataset. https://doi.org/10.17864/1947.000456 Related publication: Challen, B., Morris, M. and Cramer, R. (2023) Ultra-high-throughput and low-volume analysis of intact proteins with LAP-MALDI MS. Journal of the American Society for Mass Spectrometry, 34 (6). pp. 991-994. ISSN 1879-1123. https://doi.org/10.1021/jasms.3c00068 Contributors: Michael Morris; Waters Corporation Contact: Prof. Rainer Cramer, Department of Chemistry, University of Reading, Reading RG6 6DX, United Kingdom, Tel: 0118 378 4550, Email: r.k.cramer@reading.ac.uk.] 2. TERMS OF USE ------------ Copyright 2023 University of Reading. This dataset is licensed under a Creative Commons Attribution 4.0 International Licence: https://creativecommons.org/licenses/by/4.0/. 3. PROJECT AND FUNDING INFORMATION ------------ Title: Novel applications of liquid MALDI MS for the analysis of large biomolecules. PhD project. Dates: 2020-2023 Funding organisation: University of Reading, Waters Corporation 4. CONTENTS ------------ File listing: Figure_1.7z Figure_2.7z Figure_3.7z The files are divided into the figure they were used for. Figure_1.7z contains a .RAW and SONAR (_flatten) processed .RAW for a mixed protein solution of ubiquitin, cytochrome C and myoglobin at 17 uM, run at 10 samples/second. Figure_2.7z contains the .RAW files from Figure_1.7z, along with a .xlsx file with extracted ion currents and deconvoluted peak areas for each protein for coefficient of variation calculations. All data in the .xlsx file was taken from the SONAR processed (_flatten) .RAW file. Figure_3.7z contains 3 .RAW files - the mixed protein solution at 17 uM and at 1.7 uM, and ubiquitin at 0.17 uM. It also contains 3 SONAR (_flatten) processed .RAW files of the relevant files in the folder. All data was obtained at a rate of 10 samples/second Raw files are in a proprietary format which can be opened with the mass spectrometry vendor's software (MassLynx by Waters). All data files contain a _extern file with relevant instrument settings recorded. These can be opened in Notepad or a similar text editor. Raw data acquired with the SONAR modifications was converted to MassLynx-readable files (XX-flatten.raw) using CDTFlatten02 (custom software). Spectra were extracted manually from these datasets by combining peaks from the total ion chromatograms (TIC). Spectra extracted from single TIC peaks were deconvoluted using UniDec version 4.4.1, available as freeware from https://github.com/michaelmarty/UniDec/releases 5. METHODS ----------- The dataset contains data that was generated using a commercial mass spectrometer Synapt G2-Si HDMS (Waters) equipped with an in-house developed AP-MALDI ion source. Masslynx (ver. 4.2; Waters) software was used to collect and process the data. Further processing making use of a custom application developed by collaborators at Waters Corporation [which can be obtained on request]