Research output: Contribution to Journal/Magazine › Journal article › peer-review
Research output: Contribution to Journal/Magazine › Journal article › peer-review
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TY - JOUR
T1 - Utilization of a multiple antigenic peptide as a calibration standard in the BAN50 single antibody sandwich ELISA for Aß oligomers
AU - Kasai, Takashi
AU - Tokuda, Takahiko
AU - Taylor, Mark
AU - Nakagawa, Masanori
AU - Allsop, David
PY - 2012/6/8
Y1 - 2012/6/8
N2 - 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.
AB - 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.
KW - Amyloid-β
KW - Alzheimer’s disease
KW - Oligomer
KW - Multiple antigenic peptide
KW - ELISA
U2 - 10.1016/j.bbrc.2012.04.146
DO - 10.1016/j.bbrc.2012.04.146
M3 - Journal article
VL - 422
SP - 375
EP - 380
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
SN - 0006-291X
IS - 3
ER -