Specificity and sensitivity of the Abeta oligomer ELISA

Klaver, Andrea C.; Patrias, Lynnae M.; Finke, John M.; Loeffler, David A.

doi:10.1016/j.jneumeth.2010.12.001

Cited by 32 publications

(29 citation statements)

References 34 publications

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“…2, 3, and 7). Quite recently, the specificity of the A␤ oligomer ELISA, especially with the use of the same antibody (6E10 raised against A␤1-16) for capture and detection, has been questioned (67). In this study the use of anti-the N-terminal A␤ antibody (82E1) may help decrease the problem using the same antibodies for capture and detection, as commented by Selkoe and co-workers (38).…”

Section: Discussionmentioning

confidence: 78%

SOD1 (Copper/Zinc Superoxide Dismutase) Deficiency Drives Amyloid β Protein Oligomerization and Memory Loss in Mouse Model of Alzheimer Disease

Murakami

Murata

Noda

et al. 2011

Journal of Biological Chemistry

222

179

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Oxidative stress is closely linked to the pathogenesis of neurodegeneration. Soluble amyloid ␤ (A␤) oligomers cause cognitive impairment and synaptic dysfunction in Alzheimer disease (AD). However, the relationship between oligomers, oxidative stress, and their localization during disease progression is uncertain. Our previous study demonstrated that mice deficient in cytoplasmic copper/zinc superoxide dismutase (CuZn-SOD, SOD1) have features of drusen formation, a hallmark of agerelated macular degeneration (Imamura, Y., Noda, S., Hashizume, K., Shinoda, K., Yamaguchi, M., Uchiyama, S., Shimizu, T., Mizushima, Y., Shirasawa, T., and Tsubota, K. (2006) Proc. Natl. Acad. Sci. U.S.A. 103, 11282-11287). Amyloid assembly has been implicated as a common mechanism of plaque and drusen formation. Here, we show that Sod1 deficiency in an amyloid precursor protein-overexpressing mouse model (AD mouse, Tg2576) accelerated A␤ oligomerization and memory impairment as compared with control AD mouse and that these phenomena were basically mediated by oxidative damage. The increased plaque and neuronal inflammation were accompanied by the generation of N ⑀ -carboxymethyl lysine in advanced glycation end products, a rapid marker of oxidative damage, induced by Sod1 gene-dependent reduction. The Sod1 deletion also caused Tau phosphorylation and the lower levels of synaptophysin. Furthermore, the levels of SOD1 were significantly decreased in human AD patients rather than non-AD agematched individuals, but mitochondrial SOD (Mn-SOD, SOD2) and extracellular SOD (CuZn-SOD, SOD3) were not. These findings suggest that cytoplasmic superoxide radical plays a critical role in the pathogenesis of AD. Activation of Sod1 may be a therapeutic strategy for the inhibition of AD progression. Alzheimer disease (AD)3 is characterized by amyloid deposits in senile plaques mainly consisting of 40-and 42-mer amyloid ␤ proteins (A␤40 and A␤42) (1, 2). These proteins are produced from amyloid precursor protein (APP) by ␤-and ␥-secretases. A␤42 plays a more important role in the pathogenesis of AD than A␤40 because of its greater aggregation propensity and higher neurotoxicity (3). It has been well demonstrated that oxidative stress is a contributing factor to neurodegenerative disease progression (4, 5). A␤-induced neurotoxicity has been linked to oxidative stress via protein radicalization in vitro (6, 7). Soluble oligomeric assemblies (50ϳ60 kDa; e.g. A␤-derived diffusible ligand, A␤ * 56, and globulomer) of A␤ rather than insoluble fibrils are believed to inhibit long term potentiation and induce neuronal loss (8, 9).Many defensive systems protect mammals from oxidative stress caused by reactive oxygen species, including superoxide radicals, hydrogen peroxide, hydroxyl radicals, and singlet oxygen. Superoxide dismutase (SOD) is one of the major antioxidant enzymes that catalyzes the conversion of superoxide radicals to hydrogen peroxide (10). SOD consists of three isozymes: copper/zinc SOD (CuZn-SOD, SOD1), which is localized in the cytosol, nucle...

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Section: Discussionmentioning

confidence: 78%

SOD1 (Copper/Zinc Superoxide Dismutase) Deficiency Drives Amyloid β Protein Oligomerization and Memory Loss in Mouse Model of Alzheimer Disease

Murakami

Murata

Noda

et al. 2011

Journal of Biological Chemistry

222

179

View full text Add to dashboard Cite

show abstract

“…The majority of immunoassays currently developed to detect A␤ oligomers rely on pairs of antibodies directed against similar or identical epitopes within the A␤ peptide, thus favoring the detection of multimeric assemblies (15,80,81). Despite low detection limits, these assays cannot effectively distinguish between various forms of A␤o because the selectivity for oligomers is determined simply by number of monomers Ͼ1.…”

Section: Discussionmentioning

confidence: 99%

Prion-Protein-interacting Amyloid-β Oligomers of High Molecular Weight Are Tightly Correlated with Memory Impairment in Multiple Alzheimer Mouse Models

Kostylev¹,

Kaufman²,

Nygaard

et al. 2015

Journal of Biological Chemistry

116

132

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Background: Amyloid-␤ (A␤) oligomers are key in Alzheimer disease (AD) but are diverse and poorly characterized. Results: Multiple A␤ forms were measured across the life span of AD model mice and human AD brain. Conclusion: A␤ species interacting with prion protein were tightly linked to behavioral impairment. Significance: An A␤ oligomer subset with defined biochemical properties is present in multiple AD-relevant samples.

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“…Aβ monomer was prepared as described previously [20,21,30]. Aβ1-42 (0.5 mg; AnaSpec, San Jose, CA) was disaggregated by resuspending in 0.25 ml trifluoroacetic acid (TFA, Sigma-Aldrich, Inc., St. Louis, MO) followed by hexafluoro-2-propanol (HFIP, Sigma-Aldrich).…”

Section: Methodsmentioning

confidence: 99%

“…Aβ oligomers were also produced as described previously [20,30]. 4.8 μl of 1% NH 4 OH (AnaSpec) was added to an eppitube of disaggregated Aβ, and after brief vortexing, the tube sat for one min.…”

Section: Methodsmentioning

confidence: 99%

ELISA measurement of specific non-antigen-bound antibodies to Aβ1-42 monomer and soluble oligomers in sera from Alzheimer's disease, mild cognitively impaired, and noncognitively impaired subjects

Klaver

Coffey

Smith

et al. 2011

J Neuroinflammation

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BackgroundThe literature contains conflicting results regarding the status of serum anti-Aβ antibody concentrations in Alzheimer's disease (AD). Reduced levels of these antibodies have been suggested to contribute to the development of this disorder. The conflicting results may be due to polyvalent antibodies, antibody "masking" due to Aβ binding, methodological differences, and/or small sample sizes. The objectives of this pilot study were to compare serum anti-Aβ antibody concentrations between AD, mild cognitive impairment (MCI), and elderly noncognitively impaired (NCI) subjects while addressing these issues, and to perform power analyses to determine appropriate group sizes for future studies employing this approach.MethodsSerum antibodies to Aβ1-42 monomer and soluble oligomers in AD, MCI, and NCI subjects (10/group) were measured by ELISA, subtracting polyvalent antibody binding and dissociating antibody-antigen complexes. Differences in mean antibody levels were assessed for significance with repeated measures ANOVA using restricted maximum likelihood estimation, using Tukey-Kramer tests and confidence intervals for multiple comparisons. Spearman's rank correlation was used to determine associations between anti-monomer and anti-oligomer antibody concentrations. Estimated sample sizes required to detect effects of various sizes were calculated.ResultsThere were no significant differences between groups for mean anti-Aβ antibody levels, although these tended to be higher in AD than NCI specimens. Estimated group sizes of 328 and 150 for anti-Aβ monomer and oligomer antibodies, respectively, would have been required for 80% power for significance at 0.05 for a 25% increase in the AD mean relative to the NCI mean. Serum antibody concentrations to Aβ monomer and oligomers were strongly associated (correlations: 0.798 for undissociated sera, 0.564 for dissociated sera). Antibody-antigen dissociation significantly increased anti-Aβ monomer but not anti-Aβ oligomer antibody levels.ConclusionsThe findings in this pilot study are consistent with relatively similar concentrations of specific, non-antigen-bound antibodies to Aβ1-42 monomer and soluble oligomers in AD, MCI, and NCI sera. The differences between groups for these antibodies would have required approximate group sizes of 328 and 150, respectively, for a high probability for statistical significance. These findings do not support the hypothesis that reduced levels of anti-Aβ antibodies might contribute to AD's pathogenesis.

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Specificity and sensitivity of the Abeta oligomer ELISA

Cited by 32 publications

References 34 publications

SOD1 (Copper/Zinc Superoxide Dismutase) Deficiency Drives Amyloid β Protein Oligomerization and Memory Loss in Mouse Model of Alzheimer Disease

SOD1 (Copper/Zinc Superoxide Dismutase) Deficiency Drives Amyloid β Protein Oligomerization and Memory Loss in Mouse Model of Alzheimer Disease

Prion-Protein-interacting Amyloid-β Oligomers of High Molecular Weight Are Tightly Correlated with Memory Impairment in Multiple Alzheimer Mouse Models

ELISA measurement of specific non-antigen-bound antibodies to Aβ1-42 monomer and soluble oligomers in sera from Alzheimer's disease, mild cognitively impaired, and noncognitively impaired subjects

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