SUPPORTING INFORMATION

Electrografted Diazonium Salt Layers for

Antifouling on the Surface of SPR Biosensors

Qiongjing Zou and Karl S. Booksh*

Department of Chemistry and Biochemistry, University of Delaware, Newark, Delaware 19716,
United States
*Corresponding author: Phone: +1 (302) 831-2561. Fax: +1 (302) 831-6335. E-mail:
kbooksh@udel.edu.

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Table of contents:
Detailed Procedure of Preparation Gold-Coated Slides.
Detailed Procedure of Spectroscopic Measurements on an Electrochemical SPR Instrument.
Detailed Procedure of Preparation Diazonium Salts Solutions.
Scheme S-1. Placement of tri-pad SPR sensor in a channel.
Figure S-1. SPR wavelength as functions of refractive index of the NaCl standard solutions.

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Detailed Procedure of Preparation Gold-Coated Slides.

Briefly, BK7 glass slides (25 mm 75 mm 1 mm) for SPR analysis were cleaned by
immersion in boiling piranha solution [3:1 (v/v) concentrated sulfuric acid: 30% hydrogen
peroxide] for at least 60 min (Caution: piranha solution is strongly oxidizing and should be
handled with great care!), followed by thorough rinsing with deionized water and drying with
nitrogen gas. For tri-pad gold slides, two Scotch tape strips 0.5 mm wide each were pressed onto
the middle of the slide lengthwise. The slides were then immediately placed in a Cressington
Model 308R DC magnetron sputtering system and a 5 nm chromium adhesion layer followed by
50 nm of gold were deposited. The tape strips were then peeled off to leave two clear lines to
form electrically disconnected three gold pads on the same slide. The gold-coated slides were
used as SPR sensors immediately following deposition.

Detailed Procedure of Spectroscopic Measurements on an Electrochemical SPR

Instrument.
Briefly, the light from a white light-emitting diode (LED) source was focused in a 200 mdiameter optic fiber. Light was then collimated using a collimating lens, passed through a spatial
filter and a linear polarizer and the dove prism (BK7) with the SPR sensor contacted using
refractive index matching oil (RI = 1.5150), and collected with one or three 200 m-diameter
optic fibers depending on single-pad or tri-pad SPR sensor employed. Then, the light entered an
Andor Technology Shamrock 303 imaging spectrophotometer utilizing a thermoelectric (TE)
cooled and charge coupled device (CCD) camera. Depending on the refractive index range
desired, the spectrophotometer covered the spectral range to 59 nm with a 1200 g/mm grating or
569 nm with a 150 g/mm grating. An in-house constructed polyetheretherketone (PEEK)

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electrochemical flow-cell with an internal volume of ~180 L has two working electrodes, each
at one side, a Pt counter electrode, and a chloridized silver quasi reference electrode. Each
working electrode has one end with a spring pin, which can be pressed and connected onto SPR
gold pad, and another end outside to connect to the potentiostat. Both counter electrode and
reference electrode has one end immersed in the chamber, and another end outside to connect to
the potentiostat. When counter, reference electrodes and only one working electrode connected
to the potentiostat (Bioanalytical Systems), the electrochemical flow-cell was able to apply
potential on single-pad SPR sensor or individually on one of the outer pads of the tri-pad SPR
sensor and leave the middle pad with no potential affected.

Detailed Procedure of Preparation Diazonium Salts Solutions.

Briefly, a stock 0.005 M solution of the diazonium salt precursor was created in 0.5 M
hydrochloric acid. The diazonium salt solution was generated by adding a 50 L aliquot of 0.5 M
NaNO2 to 5.0 mL of the precursor solution (sparged with dry nitrogen gas for at least 15 min)
and allowed to react for 15 min at room temperature, in order to generate the necessary aryl
diazonium function. When 4-NDT was used, it was dissolved in 0.5 M hydrochloric acid to form
a 0.005 M solution, which was sparged with dry nitrogen gas for at least 15 min before using.

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Scheme S-1. Placement of tri-pad SPR sensor in a channel.

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Figure S-1. SPR wavelength as functions of refractive index of the NaCl standard solutions. The
line shows 2nd polynomial fit of SPR wavelength (SPR) and refractive index (); m1=18682,
m2=48320, b=31834, and R2=0.9999. The error bars depict standard deviations of three
measurements from three individual samples.

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