Conference paper
The use of proteomics for the investigation of immunostimulators and immunomodulators
Proteomics is defined as the systematic analysis of proteins expressed by an organism at a given time under certain conditions. The advantage of proteomics is that it enables investigations of numerous proteins in a single study, making it excellent for large scale screening studies. Recently the introduction of new methods has extended the field now making it well suited for hypothesis driven studies.
From a study on muscle tissue from rainbow trout (Oncorhynchuss mykiss) using a 2-dimensional gel electrophoresis (2-DE) system plentiful changes in the protein expression were found following wounding. This demonstrated the usability of proteomics to follow complicated physiological changes and responses, as several proteins were found to change in response to the wounding.
Proteins of interest to the wound healing process were then identified using Matrix-assisted laser desorption/ionization (MALDI) - mass spectrometer (MS/MS). Using the proteomic technique it was on a protein level demonstrated that the response to wounding included changes in abundance of proteins involved in cytoskeletal regulation, the immune system and iron binding capabilities.
While 2-DE is an open discovery technique meaning that no prior knowledge is required, new mass spectrometry based methods are being introduced, which require prior information on protein sequence. These new methods are excellent for hypothesis driven studies, where a large number of proteins of interest can be investigated at the same time.
Targeted mass spectrometry is such a method that allows absolute quantification (e.g. micro gram per gram of tissue) of specific proteins directly within a complex sample. When fully implemented the method can accurately quantify ~ 1000 proteins of choice with a detection limit of a few copies/cell within a single experiment.
Currently mass spectrometry based proteomics have only been set to limited use within the field of fish immunology. However these methods represent an excellent choice to supplement gene expression data and their ability to detect even low abundant proteins enable studies on proteins, which previously have been impossible by traditional approaches.
Language: | English |
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Year: | 2011 |
Proceedings: | 15th International Conference on Diseases of Fish and Shellfish |
Types: | Conference paper |
ORCIDs: | Wulff, Tune and Nielsen, Michael Engelbrecht |