Supporting Information

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1 Supporting Information Janus-Dendrimer Mediated Formation of Crystalline Virus Assemblies Joona Mikkilä, Henna Rosilo, Sami Nummelin, Jani Seitsonen, Janne Ruokolainen, Mauri A. Kostiainen*, Molecular Materials, Department of Applied Physics, Aalto University, 76 Espoo, Finland. Biohybrid Materials, Department of Biotechnology and Chemical Technology, Aalto University, 76 Espoo, Finland. Table of Contents Materials... S2 Synthesis and characterization of amphiphilic Janus-dendrimers... S2 Dynamic light scattering... S12 Gel electrophoretic mobility shift assay... S14 Small angle x-ray scattering... S15 Cryogenic transmission electron microscopy... S18 Supporting information references... S19 1 H- 1 H COSY NMR spectra of compounds 4, 5, 6 and 8... S2 S1

2 Materials All reagents were commercially available and used without any further purification. CCMV was prepared as reported in a previous studies. 1 3 The concentration of the CCMV stock solution was 1 mg L -1 in an acetate buffer (1 mm NaAc, 1 mm EDTA, 1 mm NaN 3, ph 5). In all procedures Milli-Q purified water was used. Synthesis and characterization of amphiphilic Janus-dendrimers Compounds 1 4, 7 5, 2C 12, 6 3C 6 12 and 6C 7 12 (Scheme S1) were synthesized using published procedures and all characterization data was in agreement with the literature. Column chromatography was carried out using silica gel 6 (7-23 mesh ASTM) from Fluka. Gel permeation chromatography was performed on Bio-Beads SX-1 (9:1 dichloromethane/methanol). Thin layer chromatography (TLC) was performed with Supelco aluminum-backed plates coated with.2 mm silica. Spots were visualized with ninhydrin stain (1.5 g ninhydrid, 1 ml n-butanol and 3 ml acetic acid) or by UV. The 1 H NMR and 1 H- 1 H COSY NMR spectra were recorded on a 4 MHz Bruker spectrometer. Chemical shifts are reported in parts per million (ppm) downfield of tetramethylsilane and calibrated using the residual solvent peak as an internal reference. Coupling constants (J) are reported in hertz. 1 H- 1 H COSY NMR spectra of compounds 4, 5, 6 and 8 are presented in the end of the Supporting Information. HR-ESI-MS mass spectra were recorded using Waters Micromass LCT Premier mass spectrometer. S2

3 Scheme S1. The synthesis of amphiphilic dendrimers. S3

4 2 Compound 1 (122 mg, 2.18 mmol) was dissolved in tetrahydrofuran (THF) (2 ml) and placed into an ice bath. Hydroxybenzotriazole (HOBt) (29 mg, 2.18 mmol), triethylamine (Et 3 N) (66 mg, 6.54 mmol) and N,N'-Dicyclohexylcarbodiimide (DCC) (16 mg, 7.63 mmol) were added to the solution. 7 (35 mg, 6.98 mmol) was dissolved in THF (4 ml) and added dropwise to the reaction mixture. The reaction mixture was stirred for 48 h and the temperature was allowed to rise to RT. Dicyclohexylurea (DCU) was removed by filtering twice and the solvent was evaporated under reduced pressure. The crude product was purified by silica column chromatography employing gradient elution, starting with dichloromethane/methanol (DCM/MeOH 98:2), through (9:1) to a final eluent mixture of DCM/MeOH/ammonium hydroxide (NH 4 OH) (7:1:1). The product was further purified twice by preparative gel permeation chromatography (Biobeads, DCM/MeOH 9:1) yielding product 2 (138 mg, 31 %) as a clear yellow oil. R f.53 (DCM/MeOH/NH 4 OH 7:1:1, ninhydrid stain). 1 H NMR (CDCl 3, 4 MHz) δ (81H, (CH 3 ) 3 C: 12H, CH 2 CH 2 N); 1.64 (12H, CH 2 CH 2 N); 2.41 (6H, OCH 2 CH 2 CO); (36H, CH 2 CH 2 CH 2 N); (6H, OCH 2 CH 2 CO: 6H, CCH 2 O); 4.26 (1H, Fmoc-CH); 4.33 (2H, Fmoc- CH 2 ); 7.29 (2H, aromatic Fmoc-H); 7.38 (2H, aromatic Fmoc-H); 7.61 (2H, aromatic Fmoc-H); 7.74 (2H, aromatic Fmoc-H). HR-ESI-MS (m/z): Calc. value for C 13 H 177 N 13 O 26 + Na ; found [M+Na] + ( 5. ppm). S4

5 3 Compound 2 (115 mg,.57 mmol) was dissolved in dimethylformamide (DMF) (3 ml) and piperidine (5252 mg, mmol) was added to the solution. The mixture was stirred at RT for 21 h. The solvent was evaporated and the residue was purified by gel permeation chromatography (DCM/MeOH 95:5). A yellow clear wax was recovered. Yield 997 mg (97 %). R f.18 (DCM/MeOH 93:7, ninhydrid stain). 1 H NMR (CDCl 3, 4 MHz) δ (81H, C(CH 3 ) 3 : 12H, NHCH 2 CH 2 CH 2 CH 2 NH); 1.64 (br, 12H, NCH 2 CH 2 CH 2 N); 2.41 (br, 6H, OCH 2 CH 2 CO); (36H, CH 2 CH 2 CH 2 N: 6H, CCH 2 O); 3.68 (t, 6H, J = 5.8 Hz, OCH 2 CH 2 CO). HR-ESI-MS (m/z): Calc. value for C 88 H 167 N 13 O 24 + Na ; found [M+Na] + ( 1.8 ppm). 4 Compound 3 (15 mg,.8 mmol) was dissolved in THF (8 ml). HOBt (17 mg,.13 mmol), Et 3 N (21 mg,.13 mmol) and DCC (26 mg,.17 mmol) were added to the solution. The reaction mixture was placed into an ice bath. 3C 12 (62 mg,.9 mmol) was dissolved in THF (12 ml) and added dropwise to the reaction mixture. Reaction mixture was stirred for 47 h and the temperature was allowed to rise to RT. After 24 h additional HOBt (1 mg,.7 mmol) and DCC (1 mg,.5 mmol) were added to the reaction mixture. The solvent was evaporated under reduced pressure and the crude product was S5

6 purified by gel permeation chromatography (DCM/MeOH 9:1) and silica column chromatography (DCM/MeOH 95:5) yielding product 4 (118 mg, 57 %) as a colorless wax. R f.22 (DCM/MeOH 95:5, ninhydrid stain). 1 H NMR (CDCl 3, 4 MHz) δ.87 (t, 9H, J = 6.8 Hz, CH 3 (CH 2 ) 8 ); 1.26 (br, 48H, CH 3 (CH 2 ) 8 ); (81H, C(CH 3 ) 3 : 12H, NHCH 2 CH 2 CH 2 CH 2 NH: 6H, CH 2 CH 2 CH 2 OAr); (12H, NCH 2 CH 2 CH 2 N: 6H, CH 2 CH 2 OAr); 2.42 (br t, 6H, OCH 2 CH 2 CO); (36H, CH 2 CH 2 CH 2 N); 3.72 (t, 6H, J = 6. Hz OCH 2 CH 2 CO); 3.81 (br s, 6H, CCH 2 O); 3.95 (t, 2H, J = 6.6 Hz CH 2 OAr, 4 position); 4.1 (t, 4H, J = 6.3 Hz CH 2 OAr, 3,5 positions); 6.97 (s, 2H, ArH, 2,6 positions). HR-ESI-MS (m/z): Calc. value for C 131 H 243 N 13 O 28 + Na ; found [M+Na] + (.4 ppm). 3C 12 -G1 Compound 4 (81 mg,.3 mmol) was dissolved in MeOH (3 ml) and 3 ml of MeOH/HCl (5:1) solution was added to the reaction mixture. After the addition the reaction mixture was stirred for 26 h. Solvent was evaporated under reduced pressure and one drop of HCl and 1 ml of 1,4-dioxane were added to the white solid residue to facilitate precipitation. Lyophilization gave the product 3C 12 -G1 (62 mg, 1 %) as a white solid. Yield is calculated for the HCl salt of the product. R f. (DCM/MeOH 95:5, ninhydrid stain). 1 H NMR (D 2 O, 4 MHz) δ.8 (9H, CH 3 ); 1.18 (br, 48H, CH 3 (CH 2 ) 8 ); 1.44 (6H, CH 2 CH 2 CH 2 OAr); (6H, CH 2 CH 2 OAr: 12H, NHCH 2 CH 2 CH 2 CH 2 NH); 1.91 (br, 6H CONHCH 2 CH 2 CH 2 ); 2.12 (6H, CH 2 CH 2 NH 2 ); 2.54 (br, 6H, OCH 2 CH 2 CO); (3H, CH 2 CH 2 CH 2 N); 3.25 (br, 6H, CONHCH 2 ); 3.76 (br, 6H, OCH 2 CH 2 CO); 3.83 (br, 6H, CCH 2 O); 3.91 (br, 2H, CH 2 OAr, 4 position); 4.7 (br, 4H, CH 2 OAr, 3,5 positions); 7.12 (s, 2H, ArH, 2,6 positions). HR-ESI-MS (m/z): Calc. value for C 86 H 171 N 13 O 1 + H ; found [M+H] + (.5 ppm). S6

7 5 Compound 3 (1 mg,.6 mmol) was dissolved in THF (6 ml). HOBt (13 mg,.1 mmol), Et 3 N (14 mg,.14 mmol) and DCC (36 mg,.15 mmol) were added to the solution. The reaction mixture was placed into an ice bath. 6C 12 (1 mg,.7 mmol) was dissolved in THF (6 ml) and added dropwise to the reaction mixture. Reaction mixture was stirred for 27 h after which the temperature was allowed to rise to RT. After 24 h additional HOBt (12 mg,.9 mmol) and DCC (6 mg,.3 mmol) were added to the reaction mixture. The solvent was evaporated under reduced pressure and the crude product was purified by gel permeation chromatography (DCM/MeOH 9:1) and silica column chromatography (DCM/MeOH 95:5) yielding product 5 (124 mg, 69 %) as colorless wax. R f.4 (DCM/MeOH 95:5, ninhydrid stain). 1 H NMR (CDCl 3, 4 MHz) δ.88 (2x t, 18H, overlapped, CH 3 (CH 2 ) 8 ); 1.26 (br, 96H, CH 3 (CH 2 ) 8 ); (81H, C(CH 3 ) 3 : 12H, NHCH 2 CH 2 CH 2 CH 2 NH: 12H, CH 2 CH 2 CH 2 OAr); (12H, NCH 2 CH 2 CH 2 N: 12H, CH 2 CH 2 OAr); 2.42 (br, 6H, OCH 2 CH 2 CO); (36H, CH 2 CH 2 CH 2 N); 3.72 (t, 6H, J = 5.8 Hz OCH 2 CH 2 CO); 3.81 (br s, 6H, CCH 2 O); (12H, CH 2 OAr); 4.93 (s, 4H, ArCH 2 O); 6.62 (s, 4H, ArH, 2,6 positions G2); 6.71 (1H, ArH, 4 position G1); 7.2 (br, 2H, ArH, 2,6 positions G1). HR- ESI-MS (m/z): Calc. value for C 181 H 327 N 13 O 33 + Na ; found [M+Na] + ( 3.7 ppm). S7

8 6C 12 -G1 Compound 5 (29 mg,.1 mmol) was dissolved in MeOH (4.5 ml) and 3 ml of MeOH/HCl (5:1) solution was added to the reaction mixture. After the addition the reaction mixture was stirred for 21 h. Solvent was evaporated under reduced pressure and one drop of HCl and 1 ml of 1,4-dioxane were added to the white solid residue to facilitate precipitation. Lyophilization gave the product 6C 12 -G1 (24 mg, 1 %) as a white solid. Yield is calculated for the HCl salt of the product. R f. (DCM/MeOH 95:5, ninhydrid stain). 1 H NMR (MeOD, 4 MHz) δ.9 (2x t, 18H, overlapped, CH 3 ); 1.3 (br, 96H, CH 3 (CH 2 ) 8 ); (12H, CH 2 CH 2 CH 2 OAr); (12H, CH 2 CH 2 OAr: 12H, NHCH 2 CH 2 CH 2 CH 2 NH: 6H, CONHCH 2 CH 2 CH 2 ); 2.11 (6H, CH 2 CH 2 NH 2 ); 2.49 (6H, OCH 2 CH 2 CO); (3H, CH 2 CH 2 CH 2 N); 3.26 (6H, CONHCH 2 ); 3.74 (6H, OCH 2 CH 2 CO); 3.8 (6H, CCH 2 O); (12H, CH 2 OAr); (2x s, 4H, overlapped, ArCH 2 O); (7H, ArH). HR-ESI-MS (m/z): Calc. value for C 136 H 255 N 13 O 15 + H ; found [M+H] + ( 1.9 ppm). S8

9 6 Compound 3 (152 mg,.9 mmol) was dissolved in THF (6 ml). HOBt (34 mg,.25 mmol), Et 3 N (21 mg,.21 mmol) and DCC (38 mg,.18 mmol) were added to the solution. The reaction mixture was placed into an ice bath. 2C 12 (5 mg,.1 mmol) dissolved in THF (8 ml) and added dropwise to the reaction mixture. Reaction mixture was stirred for 65 h after which the temperature was allowed to rise to RT. After 48 h additional HOBt (15 mg,.11 mmol) and DCC (1 mg,.5 mmol) were added to the reaction mixture. The solvent was evaporated under reduced pressure and the crude product was purified by gel permeation chromatography (DCM/MeOH 9:1) and silica column chromatography (DCM/MeOH 95:5) yielding product 6 (123 mg, 64 %) as a colorless wax. R f.26 (DCM/MeOH 95:5, ninhydrid stain). 1 H NMR (CDCl 3, 4 MHz) δ.87 (t, 6H, J = 6.8 Hz, CH 3 (CH 2 ) 8 ); 1.26 (br, 32H, CH 3 (CH 2 ) 8 ); (81H, C(CH 3 ) 3 : 12H, NHCH 2 CH 2 CH 2 CH 2 NH: 4H, CH 2 CH 2 CH 2 OAr); (12H, NCH 2 CH 2 CH 2 N: 4H, CH 2 CH 2 OAr); 2.41 (br t, 6H, OCH 2 CH 2 CO); (36H, CH 2 CH 2 CH 2 N); 3.72 (t, 6H, J = 5.8 Hz OCH 2 CH 2 CO); 3.8 (br s, 6H, CCH 2 O); 3.94 (t, 4H, J = 6.6 Hz CH 2 OAr, 3,5 positions); 6.52 (br s, 1H, ArH, 4 position); 6.84 (br, 2H, ArH, 2,6 positions). HR-ESI-MS (m/z): Calc. value for C 119 H 219 N 13 O 27 + Na ; found [M+Na] + ( 1.7 ppm). S9

10 2C 12 -G1 Compound 6 (99 mg,.4 mmol) was dissolved in MeOH (3 ml) and 3 ml of MeOH/HCl (5:1) solution was added to the reaction mixture. The reaction mixture was stirred for 24 h. Solvent was evaporated under reduced pressure and one drop of HCl and 1 ml of 1,4-dioxane were added to the white solid residue to facilitate precipitation. Lyophilization gave the product 2C 12 -G1 (77 mg, 1 %) as a white solid. Yield is calculated for the HCl salt of the product. R f. (DCM/MeOH 95:5, ninhydrid stain). 1 H NMR (D 2 O, 4 MHz) δ.87 (br, 6H, CH 3 ); 1.23 (br, 32H, CH 3 (CH 2 ) 8 ); (4H, CH 2 CH 2 CH 2 OAr); 1.71 (br, 4H, CH 2 CH 2 OAr); 1.83 (br, 12H, NHCH 2 CH 2 CH 2 CH 2 NH); 1.95 (br, 6H CONHCH 2 CH 2 CH 2 ); 2.14 (6H, CH 2 CH 2 NH 2 ); 2.54 (br, 6H, OCH 2 CH 2 CO); (3H, CH 2 N); 3.3 (br, 6H, CONHCH 2 ); 3.76 (br, 6H, OCH 2 CH 2 CO); 3.82 (br, 6H, CCH 2 O); 3.97 (br, 4H, CH 2 OAr, 3,5 positions); 6.48 (br s, 1H, ArH, 4 position); 6.95 (br s, 2H, ArH, 2,6 positions). HR-ESI-MS (m/z): Calc. value for C 74 H 147 N 13 O 9 + H ; found [M+H] + ( 1. ppm). 8 Compound 7 (111 mg,.22 mmol) was dissolved in THF (6 ml). HOBt (45 mg,.33 mmol), Et 3 N (5 mg,.5 mmol) and DCC (73 mg,.35 mmol) were added to the solution. The reaction mixture was placed into an ice bath. 3C 12 (159 g,.24 mmol) was dissolved in THF (8 ml) and added dropwise to the reaction mixture. Reaction mixture was stirred for 64 h after which the temperature was allowed to S1

11 rise to RT. The solvent was evaporated under reduced pressure and the crude product was purified by gel permeation chromatography (DCM/MeOH 9:1) yielding product 8 (117 mg, 46 %) as a colorless wax. R f.96 (DCM/MeOH 95:5, ninhydrid stain). 1 H NMR (CDCl 3, 4 MHz) δ.88 (t, 9H, CH 3 (CH 2 ) 8 ); 1.26 (br, 48H, CH 3 (CH 2 ) 8 ); (27H, C(CH 3 ) 3 : 4H, NHCH 2 CH 2 CH 2 CH 2 NH: 6H, CH 2 CH 2 CH 2 OAr); (4H, NCH 2 CH 2 CH 2 N: 6H, CH 2 CH 2 OAr); (1H, CH 2 CH 2 CH 2 N); 3.72 (t, 6H, J = 6. Hz OCH 2 CH 2 CO); 3.81 (br s, 6H, CCH 2 O); 3.98 (t, 2H, J = 6.6 Hz CH 2 OAr, 4 position); 4.4 (t, 4H, J = 6.3 Hz CH 2 OAr, 3,5 positions); 7.18 (s, 2H, ArH, 2,6 positions). HR-ESI-MS (m/z): Calc. value for C 68 H 126 N 4 O 1 + Na ; found [M+Na] + ( 2.2 ppm). 3C 12 -G Compound 8 (1 mg,.9 mmol) was dissolved in MeOH (3 ml) and 3 ml of MeOH/HCl (5:1) solution was added to the reaction mixture. After the addition the reaction mixture was stirred for 23 h. Solvent was evaporated under reduced pressure and two drops of HCl and 2 ml of 1,4-dioxane were added to the white solid residue to facilitate precipitation. Lyophilization gave the product 3C 12 -G (83 mg, 1 %) as a white solid. Yield is calculated for the HCl salt of the product. R f. (DCM/MeOH 95:5, ninhydrid stain). 1 H NMR (MeOD/D 2 O 1:1, 4 MHz) δ.84 (m, 9H, CH 3 ); (48H, CH 3 (CH 2 ) 8 ); (6H, CH 2 CH 2 CH 2 OAr); (br, 6H, CH 2 CH 2 OAr); (br, 4H, NHCH 2 CH 2 CH 2 CH 2 NH); (br, 2H CONHCH 2 CH 2 CH 2 ); 2.9 (quintet, 2H, CH 2 CH 2 NH 2 ); (1H, CH 2 CH 2 CH 2 N); 3.41 (br, 2H, CONHCH 2 ); 3.86 (br, 6H, CH 2 OAr); 7.5 (s, 2H, ArH, 2,6 positions). HR-ESI-MS (m/z): Calc. value for C 53 H 12 N 4 O 4 + H ; found [M+H] + ( 2.2 ppm). S11

12 Dynamic light scattering Zetasizer nano S equipment from Malvern instruments was used in DLS measurements. 3 µl of the CCMV solution (1 mg ml -1 ) was injected into 5 µl of pure water or 15 mm NaCl solution and the solutions were mixed thoroughly and filtered (Acrodisc GHP.2 µm/13 mm). After filtration the dilute CCMV solutions were titrated with aqueous dendrimer solutions (.1 mg ml -1 ) (Figure S1) that had been filtered beforehand through cotton wool. Samples were mixed properly after every dendrimer addition. The measurements were carried out by using Plastibrand semi-micro PMMA cuvettes and the temperature was kept at 25 C during the experiments. Concentrations were corrected by taking into account the sample volume left inside the syringe filter (6 µl) and volume changes after additions of the dendrimer solutions. Our previous study showed that CCMV binding affinity of cationic dendrons increases with the dendritic generation. 8 This trend was also observed with the amphiphilic Janus-dendrimers reported here. However, in our studies the effect of the hydrophobic end to the CCMV binding efficiency was revealed when the binding in different salt concentrations were compared to each other. As sperminegroup functionalized dendrons have been shown to bind CCMV less efficiently in higher salt concentrations, 8 3C 12 -G1 and 6C 12 -G1 dendrimers were found to bind more effectively in 15 mm NaCl solution than in mm NaCl solution. At the same time, binding efficiency of 2C 12 -G1 dendrimer was lower in 15 mm NaCl than in mm NaCl even though it differs from 3C 12 -G1 only by having one less hydrophobic tail. Clearly, the binding efficiency is affected by the size of the hydrophobic end of the dendrimer. S12

13 DLS in mm NaCl concentration DLS in 15 mm NaCl concentration C 12 -G1 c (mg l -1 ) C 12 -G1 c (mg l -1 ) 5 Figure S1. DLS measurements C 12 -G1 c (mg l -1 ) C 12 -G1 c (mg l -1 ) C 12 -G1 c (mg l -1 ) C 12 -G1 c (mg l -1 ) C 12 -G c (mg l -1 ) C 12 -G c (mg l -1 ) Sec. Assembly D h (nm) Sec. Assembly D h (nm) Sec. Assembly D h (nm) Sec. Assembly D h (nm) Sec. Assembly D h (nm) Free CCMV Intensity (%) Free CCMV Intensity (%) Free CCMV Intensity (%) Free CCMV Intensity (%) Free CCMV Intensity (%) Free CCMV Intensity (%) Free CCMV Intensity (%) Free CCMV Intensity (%) Sec. Assembly D h (nm) Sec. Assembly D h (nm) Sec. Assembly D h (nm) S13

14 Gel electrophoretic mobility shift assay Gel electrophoretic mobility shift assay (EMSA) experiments were carried out with BIO-RAD Power Pac Basic equipment. Agarose gels were prepared by dissolving 1 g of agarose into 1 ml of acetate buffer (1 mm NaAc, 1 mm Acetic acid, 1 mm EDTA, ph 5) and stained with 8 µl of ethidium bromide solution (.625 mg L -1 ). The CCMV concentration was 5 mg L -1 in every sample and the dendrimer concentration varied from to 8 mg L µl of 6 X Loading Dye was used to stain the samples. From the sample solutions 15 µl volumes were pipetted into the agarose gel wells. The gels were run with a constant voltage of 1 V for 3 minutes. Formation of the CCMV dendrimer complex was studied by a EMSA. Electrostatic binding and formation of complex between CCMV (5 mg L -1 ) and all four dendrimers decreased the mobility of CCMV (Figure S2). 3C 12 -G1 was the most efficient CCMV binder and could fully prevent virus migration at dendrimer concentration of 12 mg L -1. The difference between 6C 12 -G1 and 2C 12 -G1 is relatively small and both dendrimers prevented migration at 16 mg L -1. 3C 12 -G was not able to fully prevent the migration of CCMV, even at the concentration of 8 mg L -1. Lane Dendrimer c (mg l -1 ) C 12 -G C 12 -G C 12 -G C 12 -G Figure S2. Agarose gel electrophoretic mobility shift assay of free CCMV and CCMV complexed with different quantities of 3C 12 -G1, 6C 12 -G1, 2C 12 -G1 and 3C 12 -G dendrimers. S14

15 Small angle x-ray scattering The CCMV-dendrimer complex samples were prepared by mixing 5 µl of CCMV solution (1 mg ml -1 ), 1.25 µl of NaCl solution and 3.75 µl of dendrimer solution (1 mg ml -1 ). The final NaCl concentration of the 1 µl samples varied between to 3 mm. Immediately after mixing the sample solutions were sealed inside round metal holders by closing both sides of the holder with Kapton tape. Sample thickness was approximately.9 mm. The structural periodicities were measured by using a rotating anode Bruker Microstar microfocus X-ray source (Cu Kα radiation, = 1.54 Å) with Montel collimating optics. The beam was further collimated with four sets of slits (JJ X-Ray), resulting in a beam of approximately 1 x 1 mm at the sample position. The distance between the sample and the Hi-Star 2D area detector (Bruker) was 1.59 m. One-dimensional SAXS data were obtained by azimuthally averaging the 2D scattering data. The magnitude of the scattering vector is given by q = (4π/ ) sin θ, where 2θ is the scattering angle. Theoretical scattering curves were calculated with PowderCell (W. Kraus et al. J. Appl. Cryst. 29, (1996), 31) and Scatter (S. Förster et al. J. Appl. Cryst. 43, (21), 639) softwares. SAXS results measured from 3C 12 -G1, 6C 12 -G1, 2C 12 -G1 and 3C 12 -G dendrimer-ccmv complexes in different NaCl concentrations are presented in the Figures S3-S6. According to SAXS curves CCMV-3C 12 -G1 paracrystals have the largest domain sizes compared to other CCMV-dendrimer complexes as indicated by the sharp and well resolved Bragg reflections. CCMV-2C 12 -G1 complexes cause sharper reflection peaks than the CCMV-6C 12 -G1 complexes where the observed peaks were not well resolved. Also, in the case of CCMV-2C 12 -G1 complexes, the higher order reflections were visible unlike in the scattering patterns gained from the CCMV-6C 12 -G1 complexes. These observations indicate that the CCMV-2C 12 -G1 complexes have a higher degree of crystallinity than CCMV-6C 12 -G1 complexes. S15

16 Iq 2 (a.u.) Iq 2 (a.u.) c[nacl] (mm) q (nm -1 ) Figure S3. SAXS measurements of CCMV-3C 12 -G1complexes in different salt concentrations. c[nacl] (mm) q (nm -1 ) Figure S4. SAXS measurements of CCMV-6C 12 -G1complexes in different salt concentrations. S16

17 Iq 2 (a.u.) c[nacl] (mm) q (nm -1 ) Figure S5. SAXS measurements of CCMV-2C 12 -G1 complexes in different salt concentrations. Figure S6. SAXS measurements of CCMV-3C 12 -G complexes in different salt concentrations. S17

18 Cryogenic transmission electron microscopy Samples were prepared by mixing 2 µl of CCMV solution (1 mg ml -1 ),.5 µl of, 2, 24, 4 or 5 mm NaCl solution and 1.5 µl of dendrimer solution (1 mg ml -1 ) together. The resulting 4 µl sample solutions were diluted into 4 µl with, 25, 3, 5, or 625 mm NaCl solutions. Vitrification was done with Vitrobot in a saturated water vapor environment. Sample volumes of 3 µl were placed on Quantifoil R 3.5/1 grids and the excess sample was blotted away with filter paper. Blot time and drain time were both.5 s. After blotting the grids were plunged into liquid ethane/propane (1:1) solution that was cooled with liquid nitrogen surrounding the ethane/propane vessel. Imaging was performed with JEOL JEM-32FSC equipment. Images were processed with ImageJ. Figure S7. a) TEM micrograph of Heterosigma akashiwo nuclear inclusion virus (HaNIV) inclusion body 9 adapted with permission. Copyright 22, John Wiley and Sons. b) Cryo-TEM image of CCMV-3C 12 -G1 complex formed in 3 mm NaCl solution. Scale bar is 25 nm. S18

19 Figure S8. Cryo-TEM image of a highly ordered CCMV-3C 12 -G1 complex formed in 3 mm NaCl solution. Scale bar is 1 nm. Supporting information references (1) Verduin, B. J. FEBS Lett. 1974, 45, (2) Verduin, B. J. M. J. Gen. Virol. 1978, 39, (3) M. C. Aragonès, PhD thesis, The cowpea chlorotic mottle virus as a building block in nanotechnology, Radboud University Nijmegen, Nijmegen, Netherlands, 21. (4) Kostiainen, M. A.; Kotimaa, J.; Laukkanen, M.-L.; Pavan, G. M. Chem. Eur. J. 21, 16, (5) Blagbrough, I. S.; Geall, A. J. Tetrahedron Lett. 1998, 39, (6) Ropponen, J.; Nummelin, S.; Rissanen, K. Org. Lett. 24, 6, (7) Percec, V.; Ahn, C.-H.; Cho, W.-D.; Jamieson, A. M.; Kim, J.; Leman, T.; Schmidt, M.; Gerle, M.; Möller, M.; Prokhorova, S. A.; Sheiko, S. S.; Cheng, S. Z. D.; Zhang, A.; Ungar, G.; Yeardley, D. J. P. J. Am. Chem. Soc. 1998, 12, (8) Kostiainen, M. A.; Kasyutich, O.; Cornelissen, J. J. L. M.; Nolte, R. J. M. Nat. Chem. 21, 2, (9) Lawrence, J. E.; Chan, A. M.; Suttle, C. A. J. Phycol. 21, 37, S19

20 1 H- 1 H COSY NMR spectra of compounds 4, 5, 6 and 8 1 H- 1 H COSY NMR spectra of compound 4. S2

21 1 H- 1 H COSY NMR spectra of compound 5. S21

22 1 H- 1 H COSY NMR spectra of compound 6. S22

23 1 H- 1 H COSY NMR spectra of compound 8. S23