Scalable Production of Highly Sensitive Nanosensors Based on Graphene Functionalized with a Designed G Protein-Coupled Receptor

We have developed a novel, all-electronic biosensor for opioids that consists of an engineered μ-opioid receptor protein, with high binding affinity for opioids, chemically bonded to a graphene field-effect transistor to read out ligand binding. A variant of the receptor protein that provided chemical recognition was computationally redesigned to enhance its solubility and stability in an aqueous environment. A shadow mask process was developed to fabricate arrays of hundreds of graphene transistors with average mobility of ∼1500 cm2 V–1 s–1 and yield exceeding 98%. The biosensor exhibits high sensitivity and selectivity for the target naltrexone, an opioid receptor antagonist, with a detection limit of 10 pg/mL.

[1]  Jeffery G. Saven,et al.  A Computationally Designed Water-Soluble Variant of a G-Protein-Coupled Receptor: The Human Mu Opioid Receptor , 2013, PloS one.

[2]  Eric N. Dattoli,et al.  Scalable arrays of chemical vapor sensors based on DNA-decorated graphene , 2013, Nano Research.

[3]  P. Emmerson,et al.  Binding affinity and selectivity of opioids at mu, delta and kappa receptors in monkey brain membranes. , 1994, The Journal of pharmacology and experimental therapeutics.

[4]  M. Dresselhaus,et al.  Studying disorder in graphite-based systems by Raman spectroscopy. , 2007, Physical chemistry chemical physics : PCCP.

[5]  N. Kybert,et al.  Intrinsic response of graphene vapor sensors. , 2008, Nano letters.

[6]  V. Ananthanarayanan,et al.  Homology models of mu-opioid receptor with organic and inorganic cations at conserved aspartates in the second and third transmembrane domains. , 2000, Archives of biochemistry and biophysics.

[7]  Juanxia Wu,et al.  Raman spectroscopy of graphene , 2014 .

[8]  A. T. Johnson,et al.  Optimized photolithographic fabrication process for carbon nanotube devices , 2011 .

[9]  C. Saby,et al.  Electrochemical Modification of Glassy Carbon Electrode Using Aromatic Diazonium Salts. 1. Blocking Effect of 4-Nitrophenyl and 4-Carboxyphenyl Groups , 1997 .

[10]  L. Pardo,et al.  Crystal structure of the μ-opioid receptor bound to a morphinan antagonist , 2012, Nature.

[11]  S. Carroll,et al.  Evolution of Key Cell Signaling and Adhesion Protein Families Predates Animal Origins , 2003, Science.

[12]  G. Duesberg,et al.  Reliable processing of graphene using metal etchmasks , 2011, Nanoscale research letters.

[13]  M. Collins,et al.  The effects of benzodiazepines on human opioid receptor binding and function. , 2001, Anesthesia and analgesia.

[14]  Toward quantifying the electrostatic transduction mechanism in carbon nanotube molecular sensors. , 2012, Journal of the American Chemical Society.

[15]  John P. Overington,et al.  How many drug targets are there? , 2006, Nature Reviews Drug Discovery.

[16]  Mitchell B. Lerner,et al.  Hybrids of a genetically engineered antibody and a carbon nanotube transistor for detection of prostate cancer biomarkers. , 2012, ACS nano.

[17]  H. R. Krishnamurthy,et al.  Monitoring dopants by Raman scattering in an electrochemically top-gated graphene transistor. , 2008, Nature nanotechnology.

[18]  S. Bose,et al.  Recent advances in graphene-based biosensors. , 2011, Biosensors & bioelectronics.

[19]  Cinzia Casiraghi,et al.  Probing the nature of defects in graphene by Raman spectroscopy. , 2012, Nano letters.

[20]  Jeffery G. Saven,et al.  Computational design of water-soluble analogues of the potassium channel KcsA , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[21]  E. Williams,et al.  Atomic structure of graphene on SiO2. , 2007, Nano letters.

[22]  Jing He,et al.  NMR structure and dynamics of a designed water-soluble transmembrane domain of nicotinic acetylcholine receptor. , 2012, Biochimica et biophysica acta.

[23]  J. M. Perez-Aguilar,et al.  Computational Design of Membrane Proteins , 2012, Structure.

[24]  Q. Fu,et al.  Repeated growth and bubbling transfer of graphene with millimetre-size single-crystal grains using platinum , 2012, Nature Communications.

[25]  Jiwoong Park,et al.  Transfer-free batch fabrication of single layer graphene transistors. , 2009, Nano letters.

[26]  S. Banerjee,et al.  Large-Area Synthesis of High-Quality and Uniform Graphene Films on Copper Foils , 2009, Science.

[27]  James N. Weiss The Hill equation revisited: uses and misuses , 1997, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[28]  Jong-Hyun Ahn,et al.  Wafer-scale synthesis and transfer of graphene films. , 2009, Nano letters.

[29]  M. Dresselhaus,et al.  Raman spectroscopy in graphene , 2009 .

[30]  K E Carlson,et al.  Altered ligand binding properties and enhanced stability of a constitutively active estrogen receptor: evidence that an open pocket conformation is required for ligand interaction. , 1997, Biochemistry.

[31]  Pei Tang,et al.  NMR studies of a channel protein without membranes: Structure and dynamics of water-solubilized KcsA , 2008, Proceedings of the National Academy of Sciences.

[32]  B. Hong,et al.  Biomedical applications of graphene and graphene oxide. , 2013, Accounts of chemical research.