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Utilization of a multiple antigenic peptide as a calibration standard in the BAN50 single antibody sandwich ELISA for Aß oligomers

Kasai, Takashi and Tokuda, Takahiko and Taylor, Mark and Nakagawa, Masanori and Allsop, David (2012) Utilization of a multiple antigenic peptide as a calibration standard in the BAN50 single antibody sandwich ELISA for Aß oligomers. Biochemical and Biophysical Research Communications, 422 (3). pp. 375-380. ISSN 0006-291X

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Abstract

Soluble amyloid-ß (Aß) oligomers are thought to be a cause of neurodegeneration and memory loss in Alzheimer disease (AD). We recently reported a newly developed enzyme linked immunosorbent assay (ELISA) for high molecular weight (HMW) Aß oligomers in which the same Aß monoclonal antibody, BAN50, was used for both capture and detection in a single antibody sandwich ELISA (SAS-ELISA) system. Our previous data suggest that this assay will be useful for the early diagnosis of AD, but its practical application to large-scale or longitudinal studies has been limited because of lack of a reliable calibration standard. In order to develop such a standard, we have now constructed a novel peptide using the multiple antigenic peptide (MAP) technique, where multiple epitopes of BAN50 were linked, via a spacer, to a branching lysine core. We show that the standard curve constructed from a 16-mer MAP covered the physiological range of signals obtained in the BAN50 SAS-ELISA from samples of human CSF, serum, and plasma. Furthermore, this 16-mer MAP is available in large quantities and is stable against freeze–thawing. We estimate that the signal per 1 pM of this standard corresponds to 1.54–5.0 pM of HMW Aß oligomers. This MAP approach could also be used to provide an effective calibration standard for other SAS-ELISAs.

Item Type: Article
Journal or Publication Title: Biochemical and Biophysical Research Communications
Uncontrolled Keywords: Amyloid-β ; Alzheimer’s disease ; Oligomer ; Multiple antigenic peptide ; ELISA
Subjects: Q Science > QP Physiology
Departments: Faculty of Health and Medicine > Biomedical & Life Sciences
ID Code: 56678
Deposited By: ep_importer_pure
Deposited On: 13 Aug 2012 10:34
Refereed?: Yes
Published?: Published
Last Modified: 26 Mar 2013 16:00
Identification Number:
URI: http://eprints.lancs.ac.uk/id/eprint/56678

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