SUPPLEMENTARY INFORMATION

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1 SUPPLEMENTARY INFORMATION Supplementary Information A highly sensitive and selective diagnostic assay based on virus nanoparticles (Manuscript number:nnano a) Jin-Seung Park 1, Moon Kyu Cho 2, Eun Jung Lee 1, Keum-Young Ahn 1, Kyung Eun Lee 4, Jae Hun Jung 6, Yunjung Cho 3, Sung-Sik Han 4, Young Keun Kim 5, and Jeewon Lee 1 * ( These authors contributed equally to this work; *, Corresponding authors) 1 Department of Chemical and Biological Engineering, 2 Program in Micro/Nano Systems, 3 College of Medicine, 4 School of Life Sciences and Biotechnology, 5 Department of Materials Science and Engineering, Korea University, Anam-Dong 5-1, Seongbuk-Gu, Seoul , Republic of Korea. 6 Cardiovascular Division, Department of Internal Medicine, Kangnam Sacred Heart Hospital, Hallym University Medical Center, Seoul , Republic of Korea. Supplementary Methods Manufacture and SEM analysis of Ni nanohair structure First, a 250 nm thick Ag layer was deposited on the bottom side of the template by e-beam evaporation as a working electrode. Second, the electrodeposition process was performed using an electroplating bath that allows the rotation and circulation of the solution. The rotation provides uniform nanowire length while the circulation avoids any changes in the concentration of the solution. The deposition bath was composed of nickel sulfate (NiSO 4 6H 2 O), nickel chloride (NiCl 2 6H 2 O) and boric acid (H 3 BO 3 ). Nickel sulfate was the main component during the electrodeposition while nickel chloride offered enhanced conductivity. Boric acid was added for buffering purpose. The ph of the solution was fixed at 3.1 by adding nature nanotechnology 1

2 supplementary information sulfuric acid or sodium chloride, and the temperature of the bath was kept at 55~57 C. A current density of 4.07 ma/cm 2 was applied and controlled by a dc power supply (SourceMeter 2400, Keithley Instruments, USA). The deposition time was 7000 s. After the synthesis of the Ni nanowires in the template, the top side of AAO was removed (about 10~15 µm) by chemical mechanical planarization (CMP) to get flat surface and well separated Ni nanowires with uniform length. Finally, the AAO template was selectively etched by reactive ion etching (RIE, Oxford Instruments, UK) under BCl 3 environment for 30 min resulting in 3 µm long exposed Ni nanowires i.e., nanohairs. The morphology of the Ni nanohair was analyzed by field emission scanning electron microscope (FE-SEM, Hitachi S-4300) after Pt coating during 30 sec. ELISA assay All the ELISA assay experiments in this study were conducted using the commercial ELISA troponin assay kit (Troponin I EIA, Cat. No. 25-TR1HU-E01, 96 wells, ALPCO Diagnostics, NH, USA). As per the kit protocol provided by supplier, the assay procedure can be briefly summarized as follows: 1) 100 µl of troponin I that had been properly diluted in PBS buffer or human sera (AMI patient or healthy sera) was added to the antibody-coated well in 96-well microplate provided by supplier; 2) 100 µl of Enzyme Conjugate Reagent (containing anti-troponin I antibodies conjugated with HRP enzyme) was added into each well, thoroughly mixed for 30 sec, incubated at room temperature for 90 min, and then washed 5 times with distilled water; 3) After striking the wells sharply onto absorbent paper to remove all residual water droplets, 100 µl of TMB reagent (containing substrate for HRP enzyme) was added into each well, gently mixed for 5 sec, and incubated at room 2 nature nanotechnology

3 supplementary information temperature for 20 min; 4) 100 µl of Stop Solution was added to each well to stop the enzyme reaction, and then gently mixed for 30 sec; and 5) the absorbance was measured using microplate reader (GENios, Tecan, Austria) at 420 nm. Supplementary Tables Table S1. List of AMI patient and healthy sera used in this study No. Age Gender Hospital 1 74 M 2 75 F 3 58 M 4 68 M 5 63 M 6 80 M 7 59 M 8 68 M 9 74 M F F F F M M F F M M M 21* 3 F M F F F M Kangnam Sacred Heart Hospital Korea University Medical Center [*: experienced myocardial damage due to ASD (atrial septal defect) operation with extracorporeal circulation] Healthy sera No. Age Gender Hospital 1 34 M Korea University 2 24 M Medical Center 3 28 M 4 25 F 5 45 M 6 24 M 7 27 M 8 24 F 9 31 F M F Supplementary Figures M F M F M nature nanotechnology 3 Figure S1 (a)

4 supplementary information Supplementary Figures Figure S1 (a) 5 µm (b) Average pore size: nature nanotechnology

5 supplementary information Figure S2 (a) Net Photoluminescence Increase (RFU) HBV capsid-derived chimeric nanoparticles in PBS, added to the bare surface of Ni nanohair, followed by the addition of Qdotsecondary antibodies 38 nm 19 nm 3.8 nm 0.76 nm 0.58 nm 0.48 nm 0.38 nm (b) nature nanotechnology 5

6 supplementary information Supplementary Figure Legends Fig. S1 a, SEM images of Ni nanohair structures. (The upper and lower part images correspond to the top and cross-sectional views of Ni nanohair array, respectively.) b, SEM image of PVDF membrane. Fig. S2 a, Determining the amount of HBV capsid-derived chimeric nanoparticles that are added to bare nickel surface to prevent non-specific binding of Qdotsecondary antibodies to the nickel surface. b, Schematic of the panel a experiment. 6 nature nanotechnology