Smart tissue culture: in situ monitoring of the activity of protease enzymes secreted from live cells using nanostructured photonic crystals.
Monitoring enzyme secretion in tissue culture has proved challenging because to date the activity cannot be continuously measured in situ. In this Letter, we present a solution using biopolymer loaded photonic crystals of anodized silicon. Shifts in the optical response by proteolytic degradation of the biopolymer provide label-free sensing with unprecedented low detection limits (1 pg) and calculation of kinetic parameters. The enhancement in sensitivity relative to previous photonic crystal sensors constitutes a change in the sensing paradigm because here the entire pore space is responsive to the secreted enzyme rather than just the pore walls. In situ monitoring is demonstrated by detecting secretion of matrix metalloprotease 9 from stimulated human macrophages.
Kilian
Kristopher A
KA
School of Chemistry, University of New South Wales, Sydney, New South Wales 2052, Australia.
Lai
Leo M H
LM
Magenau
Astrid
A
Cartland
Siân
S
Böcking
Till
T
Di Girolamo
Nick
N
Gal
Michael
M
Gaus
Katharina
K
Gooding
J Justin
JJ
D018929:Cell Culture Techniques; D002478:Cells, Cultured; D006801:Humans; D008264:Macrophages / Q000201:enzymology; D010447:Peptide Hydrolases / Q000032:analysis* / Q000378:metabolism; D017785:Photons; D045663:Quantum Dots
D010447:Peptide Hydrolases