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Relative quantification of proteins in cell culture system exposed to biological agents by mass spectrometry

Project number: F 2509 Institution: Federal Institute for Occupational Safety and Health (BAuA) Status: Ongoing Project Planned end: 2023-09-30

Description:

Breathing offers constant opportunities for human exposure to airborne biological agents in the workplace. Although the respiratory tract possesses many barriers that prevent the uptake of biological agents, the large surface area of the lung makes it a possible portal of entry for them. Contact with biological agents can not only cause infections, but also have sensitising (allergenic) or toxic (poisonous) effects on individuals.

According to the Biological Agents Ordinance (Biostoffverordnung, BioStoffV), the infectious potential of biological agents is reflected in their classification into risk groups. However, these classifications do not take into account the biological agents’ toxic effects, and it has not been possible for such effects to be described adequately to date. Therefore, model investigations of their health-damaging impacts on human lung cells may be a suitable method with which to ascertain the toxic characteristics of biological agents.

Proteins perform functions of all kinds in cells. Furthermore, cells respond to changes like contact with biological agents in ways that alter the functions, levels of activity, or quantities of different proteins. Alterations of these kinds can be quantified by proteomic analyses, which look at the proteome, the entirety of a cell’s proteins at a particular point in time. If comparative proteomic analyses are carried out before and after contact with biological agents, particular signatures can consequently be identified that are indicative of their toxic effects on lung cells. The observed signatures could help to identify the toxic potential of biological agents. This supports the ongoing work of the Committee on Biological Agents (Ausschuss für Biologische Arbeitsstoffe, ABAS), in particular on how their toxic potential should be assessed.

The aim of this project is therefore to establish the whole workflow for the proteomic analysis of bronchial epithelial cells. This involves the cultivation of human lung cells, with or without contact to biological agents, the extraction of the proteins from the cells, and the digestion of proteins followed by the tagging of the peptides. Once they have been separated by liquid chromatography and analysed by mass spectrometry they undergo bioinformatic evaluation.

Contact

Unit 4.7 "Biological Agents"

Phone: +49 231 9071-1971 Fax: +49 231 9071-2070

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