Rapid and ultrasensitive detection of active thrombin based on the Vmh2 hydrophobin fused to a Green Fluorescent Protein
作者： Alessandra Piscitelli , Anna Pennacchio , Paola Cicatiello , Jane Politi , Luca De Stefano
DOI: 10.1016/J.BIOS.2016.09.052
关键词:
摘要: A fusion protein designed in order to combine the fluorescence emission of Green Fluorescent Protein (GFP) with adhesion ability class I hydrophobin Vmh2 was heterologously produced yeast Pichia pastoris. The Vmh2-GFP has proven be a smart and effective tool for study self-assembling. Since two proteins were linked by specific cutting site thrombin, used as active biological element realization thrombin biosensor. When present target solution specifically hydrolyzed its cleavage sequence, consequent decrease intensity sample could observed. based assay allowed quantification detection limit 2.27aM. specificity respect other proteases granted measurement added healthy human plasma same high sensitivity 2.3aM. Further advantages developed biosensor are simplicity design preparation, low requirements terms samples, reagents time.
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