title: Development of a sensitive microarray immunoassay for the quantitative analysis of neuropeptide Y.
abstract: A direct competitive immunoassay in an antibody microarray format was developed for the sensitive detection of neuropeptide Y (NPY) and employed in the analysis of NPY in human sweat samples. This is the first demonstration that antibody microarray, as a powerful multiplex analysis tool, can be used for the sensitive determination of NPY and potentially other neuropeptides. 400 pg/mL of dibiotinylated NPY and 0.1 mg/mL spotting capture antibody were found to offer the best performance, yielding a sensitivity of 50 pg/mL and a linear dynamic range of 0.1-100 ng/mL for NPY. Evaluation of matrix effects by using artificial sweat revealed that dialysis is necessary for analyzing NPY in human sweat samples with microarray immunoassay. In a preliminary application, 50-210 pg/mL of NPY was detected in sweat samples collected with Macroduct collectors. This study indicates that antibody microarrays can be used for NPY analysis and that human sweat could be a valuable sample source for biomarker and proteomics studies, especially when noninvasive human sample collection is preferable.
authors.affiliation: Section on Neuroendocrine Immunology & Behavior, National Institute of Mental Health, National Institutes of Health, Bethesda, Maryland 20892, USA.
authors.forename: Min
authors.initials: M
authors.lastname: Jia
authors.forename: Elena
authors.initials: E
authors.lastname: Belyavskaya
authors.forename: Patricia
authors.initials: P
authors.lastname: Deuster
authors.forename: Esther M
authors.initials: EM
authors.lastname: Sternberg
mesh_terms: D000906:Antibodies / Q000276:immunology; D006801:Humans; D007118:Immunoassay; D009478:Neuropeptide Y / Q000032:analysis*; D040081:Protein Array Analysis; D013542:Sweat / Q000378:metabolism
chemical_list: D000906:Antibodies; D009478:Neuropeptide Y