The 13th International Conference on Alzheimer's & Parkinson's Diseases



Austria Centre Vienna
Vienna,
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We will be presenting our newest posters on recent work in the field of neurodegeneration, copies of which will be available at our exhibit stand.

Validation of the U-PLEX Human Alpha-Synuclein Assay For Use With Human CSF, Saliva, and Whole Blood Samples Across Multiple Sites

Objectives

An analytically validated assay for human alpha-synuclein was used in a round-robin study to assess robustness across multiple laboratory sites.

Methods

The U-PLEX Human Alpha-Synuclein Kit was developed using MSD’s MULTI-ARRAY technology and validated to confirm consistency and precision. Assay components were supplied to all sites along with a common set of samples. The samples included cerebral spinal fluid (CSF), saliva, and whole blood and were a matching set inasmuch as the fluids were derived from the same individuals. Laboratory technicians were trained in the pre-analytical handling of samples as well as the execution of the assay. Four laboratories participated including sites in the USA, Sweden, and Germany. Each sample type was analyzed in duplicate across three production lots of the kits over nine days.

Results

The human alpha-synuclein assay exhibited robustness across four sites and three kit lots. The observed variability within plates, across kit lots, and across sites is reported here.

Conclusion

MSD’s validated human alpha-synuclein assay has proven robust in measuring the protein in CSF, saliva, and whole blood samples across multiple manufactured lots and multiple laboratories. The results demonstrate that the kit is suitable for long-term studies and research involving multiple international sites. Development of this assay was supported by funding from The Michael J. Fox Foundation for Parkinson’s Research (MJFF).

 

An Ultrasensitive Assay For The Detection of Phosphorylated Tau in Human Samples

Objectives

The accumulation of Tau protein in the cerebrospinal fluid (CSF) of Alzheimer’s disease (AD) patients correlates with neurodegeneration. Tau phosphorylation is connected to Tau aggregation, a pathological hallmark of multiple neurodegenerative disorders including AD. It would be advantageous to measure total and phosphorylated Tau in serum and plasma as an alternative to CSF; however, this requires assays with higher sensitivity than those currently available. Previously, we reported an ultrasensitive S-PLEX assay detecting total Tau (Nikolenko et al., 2015). In this study, we developed an ultrasensitive assay that specifically measures Tau phosphorylated at threonine 181 (T181).

Methods

An S-PLEX immunoassay was developed using antibodies that detect Tau phosphorylated at T181. This assay was used to measure phosphorylated Tau in 200 biological samples including human serum and plasma.

Results

The S-PLEX phospho-Tau T181 assay has a dynamic range of 4 logs and a lower limit of detection of 40 fg/mL. The assay specifically measures Tau phosphorylated at T181, with less than 0.01% cross-reactivity with un-phosphorylated Tau. Phosphorylated Tau was detectable in 95% of human samples tested (CSF, plasma, serum, and urine) and in cell lysates from lung, kidney, breast, and bone marrow cell lines, with concentrations ranging from 40 to 100,000 fg/mL.

Conclusion

The MSD S-PLEX phospho-Tau T181 assay enables accurate measurement of phosphorylated Tau at low concentrations in serum and plasma and may be used to detect phosphorylated Tau in biological samples.

 

Several members of the MSD team will be in attendance for in-depth discussions on how MSD assays, which deliver highly reproducible results, can empower your neurodegeneration research. Follow the links above to register at the conference website.

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