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

Research output: Contribution to Journal/MagazineJournal articlepeer-review

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  • Takashi Kasai
  • Takahiko Tokuda
  • Mark Taylor
  • Masanori Nakagawa
  • David Allsop
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<mark>Journal publication date</mark>8/06/2012
<mark>Journal</mark>Biochemical and Biophysical Research Communications
Issue number3
Volume422
Number of pages6
Pages (from-to)375-380
Publication StatusPublished
<mark>Original language</mark>English

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.