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

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• 作者：Takashi Kasai^a ; ^b ; Takahiko Tokuda^a ; Mark Taylor^b ; Masanori Nakagawa^a ; David Allsop^b ; ^{d.allsop@lancaster.ac.uk}
• 关键词：Amyloid-尾 ; Alzheimer&rsquo ; s disease ; Oligomer ; Multiple antigenic peptide ; ELISA
• 刊名：Biochemical and Biophysical Research Communications
• 出版年：2012
• 期刊代码：159_0006291x
• 类别：bio
• 出版时间：8 June, 2012
• 卷：422
• 期：3
• 页码：375-380
• 文件大小：388 K

摘要

Soluble amyloid-尾 (A尾) 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) A尾 oligomers in which the same A尾 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 A尾 oligomers. This MAP approach could also be used to provide an effective calibration standard for other SAS-ELISAs.