Table 1.

Comparison of microplate and microfluidics assay protocols for Pompe and Fabry disorders.

	Pompe assay (GAA)		Fabry assay (GLA)
	Microplate (Duke)	Microfluidics (ALL)	Microplate (Duke)	Microfluidics (ALL)
EXT	DI water	0.1% (wt/vol) Tween20 in water	1% sodium taurocholate in water	0.1% (wt/vol) Tween20 in water
Extraction time and conditions	360 μL EXT; incubated 1 h on ice	100 μL EXT; incubated 30 min at room temperature	250 μL EXT; incubated 1 h on ice	100 μL EXT; incubated 30 min at room temperature
Substrate stock	70 mmol/L in DMSO	70 mmol/L in DMSO	700 mmol/L in DMSO	700 mmol/L in DMSO
Inhibitor stock	80 μmol/L	8 mmol/L	0.5 mol/L in water	750 μmol/L in water
Final reagent during incubation	0.7 mmol/L 4-MU-α-gluc in sodium acetate, pH 3.8, with 8 μmol/L acarbose	2.5 mmol/L 4-MU-α-gluc in sodium acetate, pH 3.8, with 6 μmol/L acarbose	3.5 mmol/L 4-MU-α-gal in citrate-phosphate buffer, pH 4.5, with 75 mmol/L GalNac	5 mmol/L 4-MU-α-gal in 40 mmol/L sodium acetate, pH 4.5, with 75 μmol/L GalNac
Incubation conditions	20 h at 37 °C	6 h at 37 °C	20 h at 37 °C	6 h at 37 °C
STB	0.15 mol/L EDTA, pH 11.3	0.2 mol/L NaHCO3, pH 10.0, 0.01% (wt/vol) Tween20	0.15 mol/L EDTA, pH 11.3	0.2 mol/L NaHCO3, pH 10.0, 0.01% (wt/vol) Tween20