Coronavirus protease activity measurement method based on fluorescence resonance energy transfer
A fluorescence resonance energy and coronavirus technology, which is applied in the field of enzyme activity detection, can solve the problem that the substrate cannot be cut, and achieve the effect of improving enzyme reaction efficiency, less workload, and low detection limit
Pending Publication Date: 2021-03-09
苏州新格诺康生物技术有限公司
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Abstract
The invention provides a coronavirus protease activity measurement method based on fluorescence resonance energy transfer. The coronavirus protease activity measurement method specifically comprises the following steps: unfreezing a coronavirus active protease sample on ice; preparing a coronavirus active protease sample solution and an FRET peptide solution by using a lysis buffer solution; adding a coronavirus active protease sample solution and an FRET peptide solution into a black half-region 96-well plate; performing mixing on a shaking table for 1 minute, sealing the experiment holes byusing a microporous plate sealing film, and performing incubating at 37 DEG C; after the temperature of the pore plate is reduced to the ambient temperature, removing the sealing film of the pore plate; reading a fluorescence intensity value when the excitation wavelength/emission wavelength is equal to 340/495nm on a luminoscope; and determining the concentration of the generated Edans labeled peptide fragment by using an Edans/Dabcyl standard curve, and calculating the enzyme specific activity by using a formula. The same experimental scheme provided by the invention can be used for detecting the activities of two virus proteases.
Application Domain
Fluorescence/phosphorescence
Technology Topic
EDANSExcitation wavelength +8
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