title: Creation of stepwise concentration gradient in picoliter droplets for parallel reactions of matrix metalloproteinase II and IX.
abstract: We present a new methodology for generating a stepwise concentration gradient in a series of microdroplets by using monolithic micro valves that act as "faucets" in micrometer-scale. A distinct concentration gradient of a substrate was generated for the determination of the kinetic parameters of two different enzymes using only 10 picoliter-scale droplets. With a single experiment on a chip, we obtained K(M) and k(cat) values of matrix metalloproteinase 2 (MMP-2) and matrix metalloproteinase 9 (MMP-9), and compared the catalytic competence of the two enzymes. The present system and method are highly suitable for applications where the reagents or samples are limited and precious.
authors: Materials Research and Education Center, Department of Mechanical Engineering, Auburn University, Alabama 36849, United States.
authors: Sachin
authors: S
authors: Jambovane
authors: Duck Jong
authors: DJ
authors: Kim
authors: Evert C
authors: EC
authors: Duin
authors: Se-Kwon
authors: SK
authors: Kim
authors: Jong Wook
authors: JW
authors: Hong
mesh_terms: D055162:Biocatalysis; D002138:Calibration; D057075:Enzyme Assays / Q000379:methods*; D007700:Kinetics; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism*; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism*; D055616:Microtechnology
chemical_list: D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9