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Precipitant

Cold Room 7- 8 o C. Centrifuge. Cross-Link. Resuspend with Cold Cross linking Solution. width. Leave for 5-24 hr. Mix. length. Batch. Precipitant. Protein. 2 m m. 2-4 days at room temperature. 3X. Resuspend. Centrifuge. Centrifuge. Resuspend. Purification.

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Precipitant

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  1. Cold Room 7- 8 oC Centrifuge Cross-Link Resuspend with Cold Cross linking Solution width Leave for 5-24 hr. Mix length Batch Precipitant Protein 2 mm 2-4 days at room temperature 3X Resuspend Centrifuge Centrifuge Resuspend Purification http://www.hysitron.com/Products/products.htm

  2. Cross-link Pd2+Loading 1 mm isopropenyl acetate benzaldehyde 4-(4-methoxycarbonylphenyl)-3-buten-2-one Separation by adsorption Smaller particles More theoretical plates A) B)

  3. (1010) B) Hexagonal CPMV (0001) A) Body Centered Cubic CPMV (100) 30 nm 30 nm 60 nm 2-8 nm - ununiform pores Johnson, J. E, et. al., J. of Ultastructure Research 1981, 74, 223-231. White, J. M.; Johnson, J. E., Virology 1980, 101, 319-324.

  4. Lysozyme protein concentration B 3 mg/mL 4 mg/mL 6 mg/mL 8 mg/mL pH A C D 3.0 F E G H 3.5 J I K L 4.0 2 mm

  5. A) B) Varying temperature Varying NaCl

  6. A) C) B) Varying pH Varying temperature Varying NaCl

  7. C B Width Width (101) Length (110) c a b 800 nm lysozyme crystals Standard tetragonal lysozyme crystals b a c Morphology X-ray Diffraction A)

  8. 1 mm pH = 5.0 pH = 6.0 pH = 5.5 A) B) C) D) Literature Wang, Q., ,et. al., Angewandte Chemie-International Edition, 2002. 41(3): p. 459-462.

  9. A) The hardness of protein crystal as a function of cross-linking degree by Hysitron triboindenter. B) Multiple load-displacement plots of cystals cross-linked in 10% glutarladehyde for 24 hours Summary of the nanomechanical tests resultsfor cross-linked lysozyme crystals

  10. 50 mm (mm) A) Pd distribution by SEM D) E) B) C) Cross-linked protein crystals Soaked protein crystals with K2PdCl4

  11. Pd(II) loaded within protein crystal 6000 5500 5000 4500 c/s 4000 3500 3000 2500 2000 S of lysozyme 1500 352 350 348 346 344 342 340 338 336 334 332 880 Binding Energy (eV) 860 840 820 800 780 S of lysozyme with Pd(II) 700 680 660 640 620 600 580 178 174 170 166 162 158 Binding Energy (eV) c/s c/s A) S2p in methionine 162.59 eV S2p in cysteine disulfides 166.88 eV Pd3d in K2PdCl4 338.2 eV C) D) B) Pd bound to sulfur containing amino acids

  12. Solid Phase Conc. (ug/Kg) Aqueous Conc. (ug/L) 500 μm crystals 6 μm crystals B) Cd(II) adsorption isotherm on two sized HEWL crystals A) Cd(II) adsorption comparison with ICP-AES

  13. Benzaldehyde 4-(4-methoxycarbonylphenyl)-3-buten-2-one Isopropenyl acetate MW 146 MW 164

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