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This lecture covers the principles and techniques of protein mass spectrometry and proteomics, including protein identification using MS/MS data, direct analysis of complex protein mixtures, liquid chromatography tandem mass spectrometry, data-dependent acquisition, top-down and middle-down proteomics, sample preparation, reduction and alkylation, protein digestion, and depletion and enrichment strategies. It discusses the challenges and applications of top-down proteomics, the importance of high resolution mass spectrometry, and the use of native mass spectrometry. The lecture also addresses the significance of data analysis, the role of native mass spectrometry in disease mechanisms, and the assessment of post-translational modifications and sequence variations.