Lumbar punctures in the L4/L5 or L3/L4 interspace were used to collect CSF samples into polypropylene tubes. Prior to analysis, all CSF samples were centrifuged, aliquoted, and frozen at −80 °C. The concentrations of analyzed proteins (GPNMB, YKL-40, Aβ1-42, Aβ1-40, Tau, and pTau181) in the CSF were measured in the Department of Neurodegeneration Diagnostics, Medical University of Bialystok, Poland.
For the assessment of concentrations of neurochemical dementia diagnostics (NDD) biomarkers, IBL kits (RE59661, RE59651, Hamburg, Germany) for Aβ1-42 and Aβ1-40 and Fujirebio kits (81572, 81574, Gent, Belgium) for Tau and pTau181 proteins were used.
The analysis of GPNMB was performed using Luminex Human Discovery assay plates, provided by R&D systems, Abingdon, UK, and a Luminex 200 analyzer (Thermo Fisher Scientific, Waltham, MA, USA) (multiplexing, multiparametric, fluorescence laser reading system on microspheres for the simultaneous determination of multiple parameters). Determination of YKL-40 was performed using an ELISA kit provided by MicroVue, Quidel, San Diego, CA, USA. According to the manufacturer’s protocols, duplicate measurements were assessed for each standard, control, and sample.
For the assessment of concentrations of neurochemical dementia diagnostics (NDD) biomarkers, IBL kits (RE59661, RE59651, Hamburg, Germany) for Aβ1-42 and Aβ1-40 and Fujirebio kits (81572, 81574, Gent, Belgium) for Tau and pTau181 proteins were used.
The analysis of GPNMB was performed using Luminex Human Discovery assay plates, provided by R&D systems, Abingdon, UK, and a Luminex 200 analyzer (Thermo Fisher Scientific, Waltham, MA, USA) (multiplexing, multiparametric, fluorescence laser reading system on microspheres for the simultaneous determination of multiple parameters). Determination of YKL-40 was performed using an ELISA kit provided by MicroVue, Quidel, San Diego, CA, USA. According to the manufacturer’s protocols, duplicate measurements were assessed for each standard, control, and sample.