Luminometric sub-nanoliter droplet-to-droplet array (LUMDA) and its application to drug screening by phase I metabolism enzymes.

Here we show the fabrication of the Luminometric Sub-nanoliter Droplet-to-droplet Array (LUMDA chip) by inkjet printing. The chip is easy to be implemented and allows for a multiplexed multi-step biochemical assay in sub-nanoliter liquid spots. This concept is here applied to the integral membrane enzyme CYP3A4, i.e. the most relevant enzymatic target for phase I drug metabolism, and to some structurally-related inhibitors.

[1]  R. Rosenfeld Nature , 2009, Otolaryngology--head and neck surgery : official journal of American Academy of Otolaryngology-Head and Neck Surgery.

[2]  Charles N. Baroud,et al.  Monitoring a reaction at submillisecond resolution in picoliter volumes. , 2011, Analytical chemistry.

[3]  Bruno Pignataro,et al.  Solution processed micro- and nano-bioarrays for multiplexed biosensing. , 2012, Analytical chemistry.

[4]  S. Fabiano,et al.  Selecting speed-dependent pathways for a programmable nanoscale texture by wet interfaces. , 2012, Chemical Society reviews.

[5]  D. Chiu,et al.  Droplets for ultrasmall-volume analysis. , 2009, Analytical chemistry.

[6]  T. Sjögren,et al.  Structural basis for ligand promiscuity in cytochrome P450 3A4 , 2006, Proceedings of the National Academy of Sciences.

[7]  R. Hartmann,et al.  CYP19 (aromatase): exploring the scaffold flexibility for novel selective inhibitors. , 2008, Bioorganic & medicinal chemistry.

[8]  Hendrik Schröder,et al.  Chemische Verfahren zur Herstellung von Proteinbiochips , 2008 .

[9]  A. Ewing,et al.  Electrochemical Analysis in Picoliter Microvials , 1997 .

[10]  Kersten S. Rabe,et al.  Engineering and assaying of cytochrome P450 biocatalysts , 2008, Analytical and bioanalytical chemistry.

[11]  Sebania Libertino,et al.  On the relationship between jetted inks and printed biopatterns: molecular-thin functional microarrays of glucose oxidase. , 2009, Langmuir : the ACS journal of surfaces and colloids.

[12]  U. Doshi,et al.  Luciferin IPA–Based Higher Throughput Human Hepatocyte Screening Assays for CYP3A4 Inhibition and Induction , 2011, Journal of biomolecular screening.

[13]  David Raunig,et al.  In vitro drug interactions of cytochrome p450: an evaluation of fluorogenic to conventional substrates. , 2003, Drug metabolism and disposition: the biological fate of chemicals.

[14]  Scott L Diamond,et al.  Printing chemical libraries on microarrays for fluid phase nanoliter reactions , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[15]  Miss A.O. Penney (b) , 1974, The New Yale Book of Quotations.

[16]  T. Arakawa,et al.  Protein precipitation and denaturation by dimethyl sulfoxide. , 2007, Biophysical chemistry.

[17]  M. Uttamchandani,et al.  Nanodroplet microarrays for high-throughput enzyme screening. , 2010, Methods in molecular biology.

[18]  B. Hasinoff,et al.  Viscosity dependence of the kinetics of the diffusion-controlled reaction of carbon monoxide with the separated alpha and beta chains of hemoglobin. , 1983, Biochemistry.

[19]  G. Strambini,et al.  Glycerol effects on protein flexibility: a tryptophan phosphorescence study. , 1993, Biophysical journal.

[20]  Thomas D. Y. Chung,et al.  A Simple Statistical Parameter for Use in Evaluation and Validation of High Throughput Screening Assays , 1999, Journal of biomolecular screening.

[21]  Janos Vörös,et al.  A microwell array platform for picoliter membrane protein assays. , 2009, Small.

[22]  B. Potsaid,et al.  Development of a Fluorescence-Based, Ultra High-Throughput Screening Platform for Nanoliter-Scale Cytochrome P450 Microarrays , 2009, Journal of biomolecular screening.

[23]  A. Yarin Drop Impact Dynamics: Splashing, Spreading, Receding, Bouncing ... , 2006 .

[24]  T. Riss,et al.  Bioluminescent assays for ADMET , 2008, Expert opinion on drug metabolism & toxicology.

[25]  Philip J. Kitson,et al.  Configurable 3D-Printed millifluidic and microfluidic 'lab on a chip' reactionware devices. , 2012, Lab on a chip.

[26]  G. G. Stokes "J." , 1890, The New Yale Book of Quotations.

[27]  Bruno Pignataro,et al.  Inkjet printing methodologies for drug screening. , 2010, Analytical chemistry.