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4 protocols using platinum wire

1

Electrochemical Characterization of Biosensors

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We performed all electrochemical interrogation of our electrodes on a CH Instrument Multipotentiostat CHI1040C (Austin, TX). We used a three-electrode setup to collect measurements in which our sensors served as the working electrode, a silver/silver chloride aqueous electrode in saturated KCl (CH Instruments) served as the reference electrode, and a platinum wire (CH Instruments) completed the cell as the counter electrode. To interrogate our electrochemical cell, we applied a square-wave potential of 25 mV amplitude between −0.1 V and −0.45 V. We collected data at several square-wave frequencies over the range 5 Hz and 1 kHz but show here only the data associated with the frequency at which the sensor’s signal gain is highest. For vancomycin, phenylalanine, and aminoglycosides sensors in PBS, these frequencies are 60 Hz, 120 Hz, and 300 Hz, respectively. All measurements were conducted at room temperature.
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2

Electrochemical Characterization of Ferrocenyl-Thiol Monolayers

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NaCl, KCl, KH2PO4, NaH2PO4, and NaOH were acquired from Fischer Scientific (NJ, USA). 6-Ferrocenyl-1-hexanethiol, ferrocenemethanol (FcMeOH), 6-mercapto-1-hexanol, 1,6-hexanedithiol, dimethylsulfoxide (DMSO), phosphate-buffered saline, trifluoroacetic acid (TFA), and tris(2-carboxyethyl)phosphine hydrochloride (TCEP) were obtained from Sigma-Aldrich (MO, USA). Maleimide-modified methylene blue was obtained from ATTO-TEC GmbH (Siegen, Germany), ethanol was obtained from Gold Shield Distributors (CA, USA), H2SO4 was obtained from EMD (USA), and 2 mm gold electrodes, fritted Ag|AgCl electrodes, and platinum wire were obtained from CH Instruments (TX, USA). Microcloth (2–7/8″), 1 μm monocrystalline diamond suspension, and 0.05 μm micropolish alumina powder were obtained from Buehler (IL, USA). All were used as received. We used for all electrochemical measurements an Ag/AgCl reference, a platinum counter electrode, and a CH Instrument 660D potentiostat.
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3

Electrochemical Synthesis of Conductive Polymer Composites

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A computer-controlled potentiostat/galvanostat PGSTAT 128N equipped with Nova 1.10 software from Eco-Chemie (Utrecht, The Netherlands) was used for electrochemical polymerization of conducting polymer and electrochemical measurements. A three-electrode system was applied for all electrochemical depositions. Ag/AgCl wire was used as a reference electrode and platinum wire as a counter electrode (CH Instruments, Austin, TX, USA). ITO electrode was used as a working electrode. The electrochemical deposition was carried out from a solution containing of 50 mM of pyrrole, 10 mM of methylene blue, and one of the doping materials: 0.1 M Lac, 0.1 M Suc, or 0.01 g/L Hep. The formed Ppy and PMB layers doped with saccharides were indicated as (Ppy-PMB)Lac, (Ppy-PMB)Suc, and (Ppy-PMB)Hep. The parameters of electrochemical polymerization were selected according to our previous research [31 (link)]. Polymerization was performed at a room temperature during 25 potential cycles in the range from −0.5 V to +1.2 V vs. Ag/AgCl, at the sweep rate of 50 mV/s and step potential of 2.44 mV.
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4

Electrochemical Biosensing Reagent Preparation

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NaCl, KCl, KH2PO4, NaH2PO4, and NaOH were acquired from Fischer Scientific (NJ). H2SO4 was purchased from EMD (USA), and 2 mm gold electrodes, fritted Ag|AgCl electrodes, and platinum wire were from CHInstruments (TX). 6-Mercapto-1-hexanol, 11-mercapto-1-undecanol, 3-mercapto-1-propanol, tris(2-carboethyl)-phosphine hydrochloride (TCEP), cocaine, and murine monoclonal anti-FLAG M2 antibody were obtained from Sigma-Aldrich (MO). Kanamycin monosulfate was purchased from GoldBio.com (MO), and ethanol was obtained from Gold Shield Distributors (CA). 2 and 7/8″ microcloth, 1 μm monocrystalline diamond suspension, and 0.05 μm micropolish alumina powder were obtained from Buehler (IL). All were used as received.
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