For the preparation of sensor chips, purified IgSF-Fc proteins and normal human IgG (Sigma-Aldrich, St. Louis, MO, USA) in PBS were spotted onto an SPRi-Biochip (HORIBA France) at 10 nL/spot using a dedicated spotter in a 96-spot format at HORIBA Ltd. (Kyoto, Japan). The spotted proteins were immobilized on the chip by amine coupling. The prepared sensor chip was inserted into the instrument according to the manufacturer's instructions.
Spri biochip
Manufactured by Horiba
Sourced in France
The SPRi-Biochips™ are a line of surface plasmon resonance imaging (SPRi) sensor chips designed for use with HORIBA's SPRi detection systems. These biochips provide a platform for performing label-free, real-time monitoring of biomolecular interactions on their surface.
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Lab products found in correlation
5 protocols using spri biochip
1
Sensor Chip Preparation for IgSF-Fc Proteins
2
Peptide Microarray-based EV Detection
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Peptide microarrays were also prepared on gold SPRi chips, purchased from Horiba Scientific SAS (SPRi-Biochip), following the same protocol described above. XelPleX instrument (Horiba Scientific SAS) was firstly calibrated with a solution of 3 mg/mL of sucrose and then 500 µL of serum sample (diluted 1:10 in running buffer) were injected on the surface of the chip with a flow rate of 10 µL/min. Subsequently, 200 µL of a mixture of anti-human CD9/CD63/CD81 antibodies (500 nM each), purchased from Ancell Corporation, were injected at 25 µL/min in order to confirm the presence of EV immobilised on the chip. EzSuite and OriginLab softwares were used to analyse the SPRi signals related to each injection.
3
SPRi and Voltammetric Techniques for DNA Interactions
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Two types of gold transducers have been used in the framework of this studies. Integrated SPRi-Biochips™ made of a high refractive index glass prism coated with gold thin film were purchased in Horiba Scientific (France). Gold disk electrodes were purchased in CH Instruments (USA). SPRi experiments were conducted with the use of SPRi-Lab+ instrument (Horiba). Comparative studies of DNA-DNA interactions were performed with the use of homemade microfluidic system, equipped with two parallel cells integrated with optical system. Chronocoulometric and voltammetric measurements were carried out with CHI 1040A potentiostat equipped with homemade voltammetric cell adapted to measurements in nitrogen atmosphere. Apart from gold working electrode, Ag/AgCl/3.0 M KCl reference electrode and gold wire as auxiliary electrode were applied in a classical three-electrode system. Details of instrumentation used for AuNPs characterization (transmission electron microscopy and UV-Vis spectrophotometry) can be found in Supplementary Material.
4
SPRi Prism Functionalization Protocol
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Before the assay, the SPRi prism (SPRi-Biochips™; Horiba) was cleaned with sulfuric acid: hydrogen peroxide (4:1) solution. After washing with water and drying with N2, the SPRi prism was immersed in a 10 mM 1-dodenacothiol (Sigma-Aldrich) ethanolic solution for 12 h. Finally, the functionalized SPRi prism was thoroughly washed with ethanol and dried with N2.
5
Dual Surface Functionalization for SPRi
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For the SPRi experiments two surfaces were studied: an uncharged hydrophilic and a hydrophobic one. The sensor surface of gold-coated prisms (Horiba SPRi Biochips) was functionalized according to a protocol described by Nault et al. [36] to obtain a dual surface functionalization permitting to compare both surface chemistries in the same experiment. One side was coated with hydrophilic polyethylene glycol thiol (SH-PEG(8), ref: PEG2010.0100, Iris Biotech), diluted in water, while the other side was coated with hydrophobic 1-hexadecanethiol (SH-C 16 , ref: 52270, Merck), diluted in toluene. The prism was functionalized in two steps: first the SH-PEG solution was applied on the selected area of the prism and spread by application of a cover slip to the targeted area to be rendered hydrophilic. After 2h, the glass slide was removed and the SH-C 16 solution was applied on the entire prism for 3h. The final surfaces were thoroughly characterized previously [37] . The water contact angle was measured (DSA10-Mk2, Krüss) on both sides and was 66.1° ± 3.1° and 103.0° ± 4.1° for the hydrophilic and hydrophobic surfaces, respectively. After functionalization, the prism was used with reproducible adsorption/desorption kinetics up to four times before being functionalized again.
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