USE OF 14-3-3 PROTEINS AND A METHOD FOR DETERMINING THE SAME IN THE FLUIDS OR TISSUES OF ORGANISMS

摘要

The aim of the invention is to provide an identification method and a quantification method for 14-3-3 proteins and their isoforms for the early diagnosis of TSE diseases, which allows the diagnosis to be carried out in living organisms. A further aim is to identify a contamination of the sample by the parallel determination of a second antigen. This is achieved by the use of the biochemical characteristics of a representative of the 14-3-3 protein family, which bind to specific amino acid motives such as X(n)-XSX(n)SXXSX-X(n) or to the motive RSXpSXP within peptides or proteins. Modified solid phases, such as for example microtitre plates, are used to determine one or several isoforms or the entire 14-3-3 protein(s) and for specific binding. Said plates are coated with an artificial or natural peptide, which contains a binding motive for 14-3-3 proteins, for example a chemically synthesised peptide with the motive CAALPKINRSApSEPSLHR. After addition of the extracts or bodily fluids to be analysed, the formed peptide 14-3-3 protein complexes are detected and quantified using tagged antibodies. The inventive application of the 14-3-3 protein family and/or individual isoforms of the 14-3-3 proteins can be used as an effect monitor or biomonitor in aquatic invertebrates of environmental influences such as polychlorinated biphenyls (PCBs), (Xeno)oestrogens or other influences. The method can also be used for the early diagnosis of TSE diseases, such as Creutzfeldt-Jakob disease (CJD) and its new form (variant) in young people (vCJD) and Bovine Spongiform Encephalopathy (BSE) or similar diseases. The invention thus provides a diagnosis tag (surrogate tag), which can be used in the living organism as a screening, confirmation or sole tag.