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Biopharmaceuticals from the MS point of view
Hubálek, Martin ; Lenčo, J.
The course aims to introduce biotherapeutics, which are becoming an increasingly important part of the pharmacotherapy of serious diseases. Mass spectrometry plays an important role for their quality control in the pharmaceutical industry. We will introduce the audience to biopharmaceuticals, which in this context are peptides, proteins, monoclonal antibodies or oligonucleotides. From the analyst's point of view, we will focus mainly on antibody drugs, looking at the possibilities of their separation and analysis by mass spectrometry, at the level of intact protein molecules, antibody subunits or peptides.
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Fundamentals of proteomics
Harant, K. ; Křenková, Alena
The first part of the course will be an introduction to proteomics with the aim to show current methods and tools used for proteomic data acquisition and evaluation. In the first part, we will discuss in abbreviated form all steps of the experimental procedure from 1) designing the experiment to be suitable for subsequent LC/MS analysis, 2) sample preparation using examples of immunoprecipitation and complex lysate, to 3) measurement options using Data Dependent or Data Independent acquisition. The course will also include a critical evaluation of the quality of the measured data. The evaluation will focus on the possibilities of data processing in freeware software such as MaxQuant and DIAnn. We will also briefly introduce you to commercial programs - Proteome Discoverer, PEAKS, Spectronaut. The second part of the course will include an introduction to statistical processing of quantitative proteomic data in Perseus software. Emphasis will be placed on understanding important statistical concepts (data distribution, t-test, ANOVA, p-value, FDR, etc.) and understanding all necessary steps of statistical analysis such as data filtering, data normalization, statistical testing, etc. There will be model data that participants will process during the course.
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EI spectra interpretation
Polášek, Miroslav
Introduction to the interpretation of mass spectra. Basic mechanisms of fragmentation reactions in electron ionization. An intensive exercise on selected examples of mass spectra of electron ionization of different classes of organic compounds, which will be carried out in the spirit of the elementary didactic principle of progression from simple to complex with maximum emphasis on the independent work of the participants.\n\n
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