Supporting Information Ultrasensitive Detection of Cancer Prognostic mirna Biomarkers Based on Surface Plasmon Enhanced Light Scattering

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1 Supporting Information Ultrasensitive Detection of Cancer Prognostic mirna Biomarkers Based on Surface Plasmon Enhanced Light Scattering Chih-Tsung Yang, 1 Mohammad Pourhassan-Moghaddam, 2 Lin Wu, 3 Ping Bai 3 and Benjamin Thierry 1 * 1 Future Industries Institute and ARC Centre of Excellence in Convergent Bio and Nano Science and Technology, University of South Australia, Mawson Lakes Campus, Mawson Lakes, South Australia 5095, Australia. 2 Department of Medical Biotechnology and Immunology Research Center, Tabriz University of Medical Sciences, Tabriz 51368, Iran. 3 Electronics and Photonics Department, Institute of High Performance Computing, Agency for Science, Technology, and Research (A*STAR), Singapore *Benjamin Thierry: benjamin.thierry@unisa.edu.au S1

2 Table of Contents S1: Title page S3: Figure S1. Synthesis and modification of 55 nm AuNPs monitored by (a) UV-Vis spectroscopy and (b) DLS. (c) SEM image of the as-synthesized 55 nm AuNPs. S4: Figure S2. SPR angular spectra of DNA capture probe for the detection and amplification of (a) target mirna-122 and (b) nontarget mirna-192. S5: Figure S3. SPR kinetic measurements for mirna-122 and mirna-192. S6: Figure S4. SPR kinetic monitoring with regards to refractometric and SP-LS schemes for different concentration of the AuNP tags. S7: Figure S5. Angular/scattered intensity spectra associated to amplification using the 55 nm mab in the SP-LS scheme at mirna-122 concentrations of (a) 10 nm, (b) 1 nm, (c) 10 pm, (d) 1 pm and (e) 100 fm. S8: Table S1. Comparison of SPR based amplification schemes for the detection of mirna. S9: Figure S6. (a) Simulated angular spectra vs AuNPs pitches for 55 nm AuNPs attached on cspr substrate Field enhancement distribution associated to the presence of AuNP (55 nm) on the sensor surface for various inter-particle distance (defined as the pitch): (b) p = 600 nm and (c) p = 1500 nm. S10: Figure S7. Calibration curve of detection of mirna-122 diluted in mirna-192 (1.0 nm) with 55 nm AuNP tags. S2

3 Figure S1. Synthesis and modification of 55 nm AuNPs monitored by (a) UV-Vis spectroscopy and (b) DLS. (c) SEM image of the as-synthesized 55 nm AuNPs. S3

4 Figure S2. SPR angular spectra of DNA capture probe for the detection and amplification of (a) target mirna-122 and (b) nontarget mirna-192. S4

5 Figure S3. SPR kinetic measurements for mirna-122 and mirna-192. S5

6 Figure S4. SPR kinetic monitoring with regards to refractometric and SP-LS schemes for different concentration of the AuNP tags. Data processing for SP-LS spectra In order to quantify the scattered intensity in the SP-LS scheme, the scattered intensity peaks versus the incident angle were normalized to the same baseline to allow for meaningful comparison. Following the normalization, the scattered intensity peak was then smoothed by Origin v.9 software with adjacent-averaging of 5 points to generate a smooth curve for quantification of the scattered intensity. The change of the scattered intensity (ΔI s ) provided in Figure 3d was obtained by subtracting the scattered intensity peak before and after the binding of the 55 nm AuNP@D5H6 mab. The detailed angular/scattered spectra for the detection of different concentration of mirna-122 are shown in Figure S5. All the scattered intensity peaks were compiled for comparison with regards to different mirna-122 concentrations in Figure 3b. S6

7 Figure S5. Angular/scattered intensity spectra associated to amplification using the 55 nm mab in the SP-LS scheme at mirna-122 concentrations of (a) 10 nm, (b) 1 nm, (c) 10 pm, (d) 1 pm and (e) 100 fm. S7

8 Table S1 Comparison of SPR based amplification schemes for the detection of mirna. SPR platform Amplification scheme mirna LOD Reference SP-LS Ab conjugated AuNP mirna fm This work GOAu-SPR GO-AuNP hybrids mirna fm Wang et al. 1 SPRi AuNP mir-23b 10 fm Fang et al. 2 SPRi Silica nanoparticles fm Zhou et al. 3 SPRi AuNP mir-16,mir pm Vaisocherová et al. 4 SPRCD Antibody mirna pm Šípová et al. 5 Biacore X Supersandwich mir-21 9 pm Ding et al. 6 Biacore X Strepavidin complex mirna pm Zhang et al. 7 S8

9 Figure S6 (a) Simulated angular spectra vs AuNPs pitches for 55 nm AuNPs attached on cspr substrate field enhancement distribution associated to the presence of AuNP (55 nm) on the sensor surface for various inter-particle distance (defined as the pitch): (b) p = 600 nm and (c) p = 1500 nm. Detection of mirna-122 in mirna-192 based on SP-LS to mimic clinical condition In order to demonstrate the feasibility for the detection of target mirna in non-target mirna, mirna-122 was diluted in mirna-192 (1.0 nm) to prepare three different concentrations (1 nm, 10 pm, 1 pm) to mimic clinical samples. The RNA-DNA hybridization signal was amplified with the 55 nm AuNP@D5H6 mab for 10 min and the calibration curve is shown in Figure S7. S9

10 Figure S7. Calibration curve for the detection of mirna-122 diluted in a high mirna-192 background (1.0 nm) using the 55 nm AuNP tags. References 1 Wang, Q.; Li, Q.; Yang, X.; Wang, K.; Du, S.; Zhang, H.; Nie, Y. Graphene oxide gold nanoparticles hybrids-based surface plasmon resonance for sensitive detection of microrna. Biosens. Bioelectron. 2016, 77, Fang, S.; Lee, H. J.; Wark, A. W.; Corn, R. M. Attomole microarray detection of micrornas by nanoparticle-amplified SPR imaging measurements of surface polyadenylation reactions. J. Am. Chem. Soc. 2006, 128, Zhou, W.-J.; Chen, Y.; Corn, R. M. Ultrasensitive microarray detection of short RNA sequences with enzymatically modified nanoparticles and surface plasmon resonance imaging measurements. Anal. Chem. 2011, 83, Vaisocherová, H.; Šípová, H.; Víšová, I.; Bocková, M.; Špringer, T.; Ermini, M. L.; Song, X.; Krejčík, Z.; Chrastinová, L.; Pastva, O. Rapid and sensitive detection of multiple micrornas in cell lysate by low-fouling surface plasmon resonance biosensor. Biosens. Bioelectron. 2015, 70, Šípová, H.; Zhang, S.; Dudley, A. e. M.; Galas, D.; Wang, K.; Homola, J. Surface plasmon resonance biosensor for rapid label-free detection of microribonucleic acid at subfemtomole level. Anal. Chem. 2010, 82, Ding, X.; Yan, Y.; Li, S.; Zhang, Y.; Cheng, W.; Cheng, Q.; Ding, S. Surface plasmon resonance biosensor for highly sensitive detection of microrna based on DNA super-sandwich assemblies and streptavidin signal amplification. Anal. Chim. S10

11 Acta 2015, 874, Zhang, D.; Yan, Y.; Cheng, W.; Zhang, W.; Li, Y.; Ju, H.; Ding, S. Streptavidin-enhanced surface plasmon resonance biosensor for highly sensitive and specific detection of microrna. Microchim. Acta 2013, 180, S11