Simultaneous observation of enzyme surface diffusion and surface reaction using microfluidic patterning of substrate surfaces.
We present a study of the simultaneous observation of protease reaction and surface diffusion as the enzyme interacts with a model substrate surface. We use micro-fluidic patterning to decorate a bovine serum albumin substrate surface with stripes of adsorbed enzyme in the absence of physical barriers. Spreading of the enzyme from the initial striped region indicates surface diffusion, while removal of the substrate provides a measure of reactivity. Microfluidic patterning provides a means to determine the relative importance of enzyme adsorption, surface diffusion, and reaction on the rate of substrate removal.
Roy
Shaunak
S
Department of Chemical Engineering, Stanford University, Stanford, California 94305-5025, USA.
Thomas
Jerry M
JM
Holmes
Elizabeth A
EA
Kellis
James T
JT
Poulose
A J
AJ
Robertson
Channing R
CR
Gast
Alice P
AP
D000818:Animals; D002417:Cattle; D004058:Diffusion; D004798:Enzymes / Q000737:chemistry* / Q000378:metabolism*; D036681:Fluorescence Recovery After Photobleaching; D046210:Microfluidic Analytical Techniques / Q000379:methods*; D012710:Serum Albumin, Bovine / Q000378:metabolism; D020860:Subtilisin / Q000378:metabolism; D013499:Surface Properties
D004798:Enzymes; D012710:Serum Albumin, Bovine; D020860:Subtilisin