Biofunctionalization by spontaneous adsorption of proteins.pptVIP

SAXS: Monitoring the adsorption of BSA unloaded 296mg/g 1116mg/g Rosenfeldt et al., Phys. Rev. E 2004, 70, 061403 Proteins adsorbed onto conventional latexes: no desorption Protein is liberated when salt concentration inside layer ? salt conc. outside Release of adsorbed protein when ionic strength is raised Wittemann et al., Z. Phys. Chem. NF, in press. Adsorption of BSA: influence of pH Wittemann et al., Phys. Chem. Chem. Phys. 2003, 5, 1671. pH important but not as decisive as ionic strength Adsorption takes place on ?wrong side“ above IEP ca cs pHa pHs Wittemann et al., Phys. Chem. Chem. Phys. 2003, 5, 1671. Biesheuvel et al., J. Phys. Chem. B 2005, 109, 4209. Protein near IEP: Charge reversal by lower pH within brush Lower pH because of confined protons protonation of NH2 groups on surface Charge reversal cannot be the only reason for the strong adsorption! But: Glucoamylase (pI = 3.5) is adsorbed at pH = 6.1 Neumann et al., Macromol. Biosci. 2004, 4, 13; Haupt et al., Biomacromolecules 2005, 6, 948. blue: basic residues red: acidic residues yellow: neutral residues Protein surface: ?patches“ of positive and negative charge Positive patches become counterions for polyelectrolyte chains of SPB - - - - - - - - - - + + + + - - - - - - * Biofunctionalization by spontaneous adsorption of proteins K. Henzler, A. Wittemann, S. Rosenfeldt, B. Haupt, M. Ballauff University of Bayreuth Aim: Immobilization of proteins on colloidal carriers ?bionanoparticles“ Colloidal particles provide large surfaces large amount of immobilized biomolecules Enzymes can be used as catalysts for technical applications substrate bound enzymes product Problem: Adsorption on solid surfaces has to be avoided! M. Santore et al., Langmuir 2002, 18 (3), 706. Adsorption on solid surface Denaturation of proteins by adsorption on solid surfaces strong attraction by van der Waals or hydrophobic interaction Loss of biological function Long charged polyelectrolytes attached