UGOTUniversity of Gothenburg (Sweden)
UGOTUpper Grand Occasional Teachers (Canada)
UGOTUrinary Glutamic-Oxaloacetic Transaminase (biochemistry)
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Neurogranin was recognized in CSF by the WashU and UGot antibodies, i.e., bands were shown, unexpectedly, around 60 or 70 kDa, but not by the ADx antibodies.
The WashU assay fully recognized the UGot calibrator, and the neurogranin values obtained by the WashU assay were approximately 30% higher than the concentrations obtained by the UGot assay itself.
The WashU and UGot assay quantified neurogranin in the brain lysate at similar concentrations, whereas the ADx assay detected approximately 70-fold lower neurogranin concentrations compared with the WashU and UGot assays.
Neurogranin concentrations could be determined in all samples by using the WashU and ADx assays, whereas 9 out of 108 samples were below the LLOQ by using the UGot assay.
Effect size was the highest in the UGot assay: [[eta].sup.2] for the WashU, ADx, and UGot assays was 0.14, 0.11, and 0.21, respectively.
A weak negative correlation between neurogranin and MMSE score was found in the UGot assay ([rho] = -0.23, P < 0.05) butnotin the WashU or ADx assays (see Material 5 in the online Data Supplement).
All assays detected different neurogranin epitopes: the WashU assay targets the N-terminal and C-terminal part of the protein, whereas the UGot ELISA specifically targets the C-terminal end.
The WashU and ADx calibrators showed clean bands at 14 and 6 kDa, respectively, and the UGot calibrator showed many protein bands at different molecular weights on silver stain, likely due to BSA that was added to the calibrator solution.
Importantly, neurogranin concentrations among all 3 assays correlated well in all clinical dementia groups (overall, [rho] = 0.95 between ADx and WashU, [rho] = 0.87 between UGot and WashU, and [rho] = 0.81 between UGot and ADx).
WashU targets full-length neurogranin, ADx targets P75-truncated neurogranin, and UGot targets neurogranin at the C-terminus.
[11] Nonstandard abbreviations: AD, Alzheimer disease; CSF, cerebrospinal fluid; WashU, Washington University; ADx, ADx Neuro Sciences; UGot, University of Gothenburg; DLB, dementia with Lewy bodies; FTD, frontotemporal dementia; VaD, Vascular dementia; PBST, PBS containing 0.05%Tween-20; LLOQ, lower limit of quantification; ANCOVA, analysis of covariance; MMSE, Mini-Mental State Examination.
The 3 rows above ("antibodies") indicate what epitopes are tagged by the capture and detection antibodies used in the WashU assay (upper row: S10-D23 and G49-G60), ADx assay (middle row: R53-A64 and G62-P75), and UGot assay (bottom row: G52-G65 and V66 -D78).