INNOTEST® PHOSPHO-TAU (181P)

Single-analyte assays using Elisa technology 

The INNOTEST PHOSPHO-TAU(181P) is a solid-phase enzyme immunoassay for the quantitative determination of phosphorylated Tau (Phospho-Tau(181P)) in human cerebrospinal fluid (CSF).

The combined use of CSF-Tau and CSF-β-amyloid(1-42) marker concentrations allows differentiation between Alzheimer’s disease (AD) and normal aging or other neurological diseases such as depression.1-5

The discrimination of AD from non-AD types of dementia such as dementia with Lewy Bodies may be further improved using the quantification of CSF-phospho-Tau(181)6-7 

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For research use only
INNOTEST® PHOSPHO-TAU (181P)

Product number 81581

96 Tests
Please contact your local Fujirebio representative for the availability of this product in your country.

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    Advantages of our assays

    • Simple colorimetric immunoassays that are easily automated on microplate-processor
    • Supported by many peer-reviewed scientific publications
    • Less than 300 μL of CSF necessary for determination of complete biomarker profile
    • Assay range consistent with biological range of values

      
    Clinical background

    Alzheimer’s disease (AD) is the most common form of dementia and is histologically characterized by the accumulation of extracellular amyloid plaques and intracellular neurofibrillary tangles throughout the brain. The major constituents of amyloid plaques are the β-amyloid peptides consisting of 40 and 42 amino acids, which are derived from the amyloid precursor protein. Neurofibrillary tangles are made up of paired helical filaments consisting of hyperphosphorylated tau protein (phospho-tau). Tau protein, present in the brain in 6 different isoforms, is an intracellular protein that is released upon neuronal death.

    A license regarding amyloid beta antibodies contained in this product under patents US 570349 and EP 0683234 has been obtained from Takeda Pharmaceutical Company Limited. Furthermore, a license for the use of amyloid beta monoclonal antibodies contained in this product under patents US 5,593,846A, US 5,766,846A, US 5,837,672A, US 6,284,221B1, US 6,610,493B1, US 5,441,870A, US 5,721,130A, US 5,605,811A and US 6,114,133A has been obtained from Eli Lilly and Company.

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    Can the INNOTEST β-Amyloid(1-42), the INNOTEST β-Amyloid(1-40), INNOTEST hTau Ag and INNOTEST PHOSPHO-Tau(181P), and INNO-BIA Plasma AΒ forms assay be used for samples from species other than humans?

    If the species has the same amino acid sequence as humans or a highly conserved amino acid sequence, particularly in the areas that are recognized by the antibodies (epitopes) in the assays, it is possible to use the CSF or plasma of other species on these assays.*

    The human amino acid sequence for β-amyloid(1-42) and Tau are listed below, together with the epitopes of our antibodies used in the Neuro assays. You can compare these with the sequence of the other species.

    A recommended site to look up amino acid sequences of other species is www.uniprot.org.

    • 81576 (CE) / 81583 (RUO): INNOTEST β-AMYLOID(1-42): antibodies used are 3D6 and 21F12.

    neurodegeneration faq 1

    • 80462 (CE) / 81585 (RUO): INNOTEST β-AMYLOID(1-40): antibodies used are 3D6 and 2G3.

    neurodegeneration faq 2

    • 81572 (CE) / 81579 (RUO): INNOTEST hTau Ag: antibodies used are AT120, HT7 and BT2
    • 81574 (CE) / 81582 (RUO): INNOTEST PHOSPHO-TAU(181P): antibodies used are AT270 and HT7

    neurodegeneration faq 3

    • 80933: INNO-BIA plasma Aβ forms: antibodies used are 3D6, 2G3 and 21F12 (cf. INNOTEST assays)

    * Use on other samples than CSF is not validated by Fujirebio Europe N.V.

    Can brain homogenates be tested on the neuro INNOTEST assays?

    Yes, below you can find a protocol and references to publications where it has been used in combination with our Neuro INNOTEST® assays.*

    Protocol for brain Aβ42 and Aβ40:

    • Homogenize mouse brain in 6.5 volumes of 20 mM Tris-HCl, pH 8.5 with
      5 mM EDTA,
      2 mM phenylmethylsulfonyl fluoride
      0.5 mg/ml leupeptin
      0.7 mg/ml pepstatin
      0.1 mg/ml phenanthroline
      0.1 mg/ml benzamidine
    • Clear the homogenate as follows: centrifuge at 135,000 x g for 1h at 4°C and at 220,000 x g for 2h at 4°C.
    • Concentrate on Sepak C18 cartridges (Waters, Milford, MA).

    This protocol has been used in the following publications:

    • Moechars et al., 1999 J Biol Chem
    • Lee et al., 2003 J Biol Chem

    * These protocols were not validated by Fujirebio Europe N.V.

    Is there a correlation between the concentrations obtained with the INNO-BIA AlzBio3 and the INNOTEST assays?

    Although there is a difference in absolute concentrations between both assays, INNOTEST and INNO-BIA AlzBio3* results are highly correlated (cf. publication list).  However, for the conversion of INNOTEST concentrations into INNO-BIA concentrations or vice versa (e.g. pooling of two different data sets), we highly recommend to define your own conversion factor by analyzing at least a subset of your study population with both technologies, given the fact that:

    • different publications mention different correlation factors and conversion factors
    • the conversion factor might not be the same for the entire measurement range
    • there is no certified reference material available to which the concentrations obtained with these assays can be linked.

    Publication list:

    • Olsson et al. Clin Chem 2005;51(2):336-345
    • Reijn et al. Clin Chem 2007;53:859-65
    • Lewczuk et al. Neurobiol Aging 2008 Jun;29(6):812-8
    • Wang et al. J Alzheimers Dis 2012;31(2):439-45
    • Irwin et al. Arch Neurol 2012 Aug ;69(8) :1018-25
    • Le Bastard et al. J Alzheimers Dis 2013;33(1):117-31
    • Vanderstichele et al. Clin Chem 2013 Apr;59(4):710-2.

    * The INNO-BIA AlzBio3 assay is no longer commercially available.

    How do I define a cut-off value?

    Cut-off values are usually calculated using samples from clinically diagnosed AD patients and healthy controls or non-AD dementia patients that have been followed up for some years.  This follow-up is important since it possibly impacts the obtained optimal cut-off value and/or its associated sensitivity and specificity, because:

    1. a significant number of patients with an AD clinical diagnosis do not have an underlying AD pathology and;
    2. AD pathology may be present decades before the onset of the clinical symptoms. 

    The samples used for setting the cut-off values should preferably have undergone the same pre-analytical track as the samples that will be analyzed and interpreted in routine, as many of these pre-analytical factors influence the final biomarker results (Vanderstichele et al. Alzheimers Dement 2012 Jan;8(1):65-73 - del Campo et al. Biomark Med 2012 Aug;6(4):419-30).

    Different statistical approaches to choosing the right cut-off are described in Bartlett et al. Biomark Med 2012. Sample sizes should be chosen so that cut-off values can be estimated with adequate precision and this will depend on the statistical approach.    

    According to the consensus report, the (combined) assessment of biomarkers for Alzheimer’s disease should achieve at least 80% sensitivity and specificity (Working Group on Molecular and Biochemical Markers of Alzheimer’s disease, Neurobiol Aging 1998). Cut-off values determined in one population should preferably be cross-validated in an independent cohort to avoid overestimation of the diagnostic performance of the chosen cut-off values (Bartlett et al. Biomark Med 2012 Aug;6(4):391-400).

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