Table 1.

An overview of the conditions optimized for the current TDP-43 assay. This table presents the optimized parameters for the Meso Scale Discovery (MSD) assay using the specified antibody pair. The parameters include antibody concentrations, incubation times, buffer compositions, and detection settings. Each parameter has been meticulously fine-tuned to ensure maximum assay sensitivity and specificity for the target analyte.

Assay Parameter	Variables Tested		Variable Selected
Antibodies	6 antibodies in all combinations (see Figure 1B)		Capture: PtG Rb 10782 Detection: R&D Ms 671818R
Blocking reagents	Casein/TBS 1% BSA in Casein/TBS Iron Horse Assay Diluent/PBS TRIPLE Block (1% Casein, 1% BSA and 1% fish gelatin) +Tween-20		1% BSA in Casein/TBS
Blocking time and temperature	2 hrs at 37°C, Shaking (750rpm) Overnight at 4°C, No Shaking 2 days in 4°C, No Shaking		2 hrs in 37°C, Shaking (750rpm)
Diluents	Casein/TBS Casein/TBS+Triton Casein/TBS+Tru Block Casein/TBS+Tru Block+Triton Casein/TBS+Tru Block+Tween-20 Iron Horse Assay Diluent/PBS		Iron Horse Assay Diluent/PBS
Sample dilutions for plasma	2-fold 4-fold 8-fold 16-fold	32-fold 64-fold 128-fold 256-fold	4-fold
Sample dilutions for serum	2-fold 4-fold 8-fold 16-fold	32-fold 64-fold 128-fold 256-fold	4-fold
Capture antibody concentrations	0.5 μg/mL 0.75 μg/mL	1.0 μg/mL 1.5 μg/mL	1.5 μg/mL
Detection antibody concentrations	0.5 μg/mL 0.75 μg/mL	1.0 μg/mL 1.5 μg/mL	0.5 μg/mL
SULFO-Tag concentrations	0.5 μg/mL 0.75 μg/mL	1.0 μg/mL 1.5 μg/mL	1.0 μg/mL