Quantitation of amyloid-beta peptides in biological milieu using a novel homogeneous time-resolved fluorescence (HTRF) assay.
Many of the recent advances in the understanding of the pathological processes underlying Alzheimer's disease have come about as a result of the development of assays that can specifically quantitate in biological milieu amyloid-beta (A beta) peptides ending at amino-acid positions Ala-42 (A beta(42)) and Val-40 (A beta(40)). The existing technologies, however, although proven in their utility are limited in their application with regards to sample manipulation and suitability for high-throughput screening. To overcome these limitations, in this report we describe the development of a novel homogeneous time-resolved fluorescence (HTRF) immunoassay for A beta(42) and A beta(40) peptides. This assay has the sensitivity, selectivity and dynamic range to allow specific, direct quantitation of A beta peptides in cell culture medium, plasma, cerebrospinal fluid and brain tissue extracts, and has the major advantage of minimising sample manipulation and its inherent inaccuracies.
Clarke
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EE
Department of Molecular Biology, Merck Sharp & Dohme Research Laboratories, Neuroscience Research Centre, Terlings Park, Harlow, Essex CM20 2QR, UK. earl_clarke@merck.com
Shearman
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MS
D000328:Adult; D000544:Alzheimer Disease / Q000097:blood / Q000134:cerebrospinal fluid / Q000175:diagnosis*; D016229:Amyloid beta-Peptides / Q000032:analysis* / Q000097:blood / Q000134:cerebrospinal fluid; D000918:Antibody Specificity / Q000276:immunology; D000936:Antigen-Antibody Complex / Q000032:analysis; D001774:Blood Chemical Analysis; D002478:Cells, Cultured / Q000276:immunology; D003470:Culture Media / Q000032:analysis; D015200:Fluoroimmunoassay; D006801:Humans; D008297:Male; D010455:Peptides / Q000032:analysis* / Q000097:blood / Q000134:cerebrospinal fluid; D017434:Protein Structure, Tertiary; D013997:Time Factors
D016229:Amyloid beta-Peptides; D000936:Antigen-Antibody Complex; D003470:Culture Media; D010455:Peptides