Table 3
Comparisons of existing sialidase assay methods
TBA assay TLC Radiochemical method 1H-NMR HPLC Fluorescence assay Spectophotometric assay
Substrates Sialosides Sialosides Sialosides Sialosides Sialosides 4MU-Neu5Ac pNP-Neu5Ac
Reagents Arsenite, TBA Resorcinol or diphenylamine Radiolabelled gangliosides Deuterated buffer Malonitrile Normal buffer Normal buffer
Instruments Spectrometer TLC plate HPTLC and radiochromatoscanner NMR facility HPLC and fluorimeter Fluorimeter Spectrophotometer
Time to run hours hours hours hours hours 10 min 10 min
Safety issue 5TC toxic Hazardous in case of skin contact Radioactive substrate Strong magnet None None None
Automation No No No No Yes Yes Yes
Cost per run Cheap Cheap Expensive Very Expensive Expensive Expensive and substrate unstable Cheap
Substrate specificity Yes Yes Yes Yes Yes No No
Enzyme kinetics Inaccurate; Time consuming Not applicable Costly; Time consuming Costly; Time consuming Costly; Time consuming Yes Yes
Reference(s) [8] [9] [10] [3,11] [20] [7,12,16] [12]

Hayre et al. BMC Biochemistry 2012 13:19   doi:10.1186/1471-2091-13-19

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