title: Development of a specific ELISA to measure BACE1 levels in human tissues.
abstract: The aspartyl protease BACE1 is the rate limiting enzyme in the synthesis of amyloid beta, which accumulation in the human brain is a hallmark of Alzheimer's disease (AD). BACE1 has been proposed as a surrogate marker of AD; however, very few BACE1 immunoassays have been reported. In the present study we have screened ten BACE1 antibodies by Western blot and several antibody pairs to develop a new BACE1 sandwich ELISA procedure. We identified one pair that showed little background and good reproducibility. Several dilution buffers and sample denaturation methods were tried to partially unfold BACE1 before capture. We found that dilution in PBS followed by 10 min incubation at 50°C critically improves the performance of the assay. Finally, we successfully measured BACE1 levels in a few human brain and platelet lysates as well as in plasma and AD CSF. We anticipate that this assay will lay the ground to accurately measure BACE1 levels in human tissues, which could facilitate the molecular diagnosis of AD in the near future.
authors: Banner Sun Health Research Institute, Haldeman Laboratory of Molecular Diagnostics and Therapeutics, 10515 Santa Fe Drive, Sun City, AZ 85351, USA. Amanda.Gonzales@bannerhealth.com
authors: Amanda
authors: A
authors: Gonzales
authors: Boris
authors: B
authors: Decourt
authors: Aaron
authors: A
authors: Walker
authors: Rachel
authors: R
authors: Condjella
authors: Hikmet
authors: H
authors: Nural
authors: Marwan N
authors: MN
authors: Sabbagh
mesh_terms: D000544:Alzheimer Disease / Q000175:diagnosis / Q000201:enzymology; D053829:Amyloid Precursor Protein Secretases / Q000032:analysis*; D000818:Animals; D000906:Antibodies; D000918:Antibody Specificity; D016282:Aspartic Acid Endopeptidases / Q000032:analysis*; D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D000906:Antibodies; D053829:Amyloid Precursor Protein Secretases; D016282:Aspartic Acid Endopeptidases; C509280:BACE1 protein, human