NR AHFJ
AU Leuck,J.; Peoch,K.; Laplanche,J.L.; Grabar,E.; Müller,W.E.G.; Schröder,H.C.
TI A novel binding assay for detection of 14-3-3 protein in cerebrospinal fluid from Creutzfeldt-Jakob patients
QU Cellular and Molecular Biology (Noisy-le-Grand, France) 2000 Nov; 46(7): 1291-6
PT journal article
AB Here we describe a simple and fast microplate assay for 14-3-3 protein, which is based on the binding of 14-3-3 protein to a chemically synthesized peptide containing the phosphorylated recognition sequence of this protein. The peptide is covalently bound to maleimide-activated microplates through reaction of the sulfhydryl group of a N-terminal cysteine residue linked to the peptide. The peptide- 14-3-.3 complexes formed are detected using - as a second recognition principle - an anti-14-3-3 protein antibody. This assay was used for measurement of 14-3-3 protein in brain (bovine) and in cerebrospinal fluid from patients with Creutzfeldt-Jakob disease (CJD) and control cases with diseases other than CJD. This novel assay allowed also the detection of low 14-3-3 protein levels present in cerebrospinal fluid, without further concentration, in contrast to conventional immunoblotting or immunoassay procedures.
MH Aged; Amino Acid Motifs; Amino Acid Sequence; Animal; Antibodies/immunology; Binding Sites; Brain Chemistry; Cattle; Creutzfeldt-Jakob Syndrome/*cerebrospinal fluid; Human; Immunoassay/*methods; Immunoblotting; Molecular Sequence Data; Peptides/chemical synthesis/chemistry/metabolism; Phosphoproteins/cerebrospinal fluid/immunology/metabolism; Phosphorylation; Protein Binding; Sensitivity and Specificity; Support, Non-U.S. Gov't; Tyrosine 3-Monooxygenase/*cerebrospinal fluid/immunology/metabolism
AD Institut für Physiologische Chemie, Universität Mainz, Germany.
SP englisch
PO Frankreich