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High Molecular Amyloid β Oligomer ELISA Kit Wako Ver.2 Technical Information and Data

Assay principal

This ELISA kit is designed to measure ≧ 9-mer Aβ oligomer specifically by using same Aβ antibody (BAN50) for capture and detection3)4). As a standard, a 16-mer MAP peptide with 16 epitopes of anti Aβ antibody (BAN50) is used4).

3) Fukumoto, H., et al.: FASEB J., 24, 2716(2010).
4) Kasai, T., et al.: Biochem. Biophys. Res. Commun., 422, 3, 375(2012).


Specificity for Aβ oligomers in the ELISA

Aβ monomers were incubated the solution at 4℃ overnight in order to synthesize various size of Aβ oligomers. They were separated Fraction A and Fraction B by ultrafiltration. The fractions were analyzed by western blotting and the Aβ oligomer ELISA.

Western blotting using Aβ antibody(BAN50)

Aβ oligomer ELISA

A strong signal was exhibited in the fraction A containing 9-mer and more (> 30 kDa), whereas few signal was detected in fraction B containing 8-mer and smaller (< 30 kDa).


Example of measurements

Example of measurement 1 - spiked recovery test and dilution linearity

Human plasma Not spiked +0.5pM Std. +0.1pM Std. +5pM Std. +30pM Std.
Assay value Assay value Recovery Assay value Recovery Assay value Recovery Assay value Recovery
1.85pM 2.36pM 102% 2.85pM 100% 7.11pM 105% 31.2pM 97.8%
Human serum Not spiked +0.5pM Std. +0.1pM Std. +5pM Std. +30pM Std.
Assay value Assay value Recovery Assay value Recovery Assay value Recovery Assay value Recovery
1.58pM 2.07pM 98% 2.59pM 101% 6.64pM 101% 31.9pM 101%

 

Human CSF1 Not spiked +3pM Std. +6pM Std. +15pM Std.
Assay value Assay value Recovery Assay value Recovery Assay value Recovery
5.16pM 8.01pM 95% 10.6pM 91% 18.1pM 86%
Human CSF2 Not spiked +3pM Std. +6pM Std. +15pM Std.
Assay value Assay value Recovery Assay value Recovery Assay value Recovery
29.1pM 38.8pM 97% 44.0pM 99.3% 59.9pM 88%

 


Example of measurement 2 - CSF sample

Cerebrospinal fluids(CSF) of patients with Alzheimer disease (AD) and Mild Cognitive Impairment (MCI), non-dementia patients(control), were measured with our ELISA kit and competitor’s kit.

Human Serum

  Age Sex MMSE※3 CSF
Aβ oligomer(pM)
Control
n=5
69 M 30 2.37
60 F 30 1.65
59 M 30 2.82
46 M 28 2.24
60 F 29 1.34
MCI
n=5
69 F 24 3.6
76 M 23 2.62
82 M 26 2.33
69 F 23 2.67
73 F 25 2.34
AD
n=8
79 F 10 3.56
88 M 24 3.13
62 M 16 4.13
78 M 13 3.95
63 M 14 3.88
74 F 16 3.25
76 M 19 3.69
84 M 16 4.34

***MMSE score:diagnosis Inquiry test score of cognitive function
30-28 Normal
27-24 Suspected of mild cognitive impairment
≦ 23 Suspected of dementia

Human Plasma

 

  • There was a significant difference between Control, MCI and AD.
  • High molecular Aβ oligomer correlated with MMSE score.
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