pH dependence and protein selectivity of poly(ethyleneimine)/poly(acrylic acid) multilayers studied by in situ ATR-FTIR spectroscopy.
The selective interaction between polyelectrolyte multilayers (PEM) consecutively adsorbed from poly(ethyleneimine) (PEI) and poly(acrylic acid) (PAC) and a binary mixture containing concanavalin A (COA) and lysozyme (LYZ) based on electrostatic interaction is reported. The composition and structure of the PEM and the uptake of proteins were analyzed by in situ attenuated total reflection (ATR) Fourier transform infrared (FTIR) spectroscopy, and the morphology and thickness were characterized by atomic force microscopy (AFM) and ellipsometry. The PEM dissociation degree and charge state and the protein adsorption were shown to be highly dependent on the outermost layer type and the pH in solution. High protein uptake was obtained under electrostatically attractive conditions. This was used to bind selectively one protein from a binary mixture of LYZ/COA. In detail it could be demonstrated that six-layered PEM-6 at pH = 7.3 showed a preferential sorption of positively charged LYZ, while at PEM-5 and pH = 7.3 negatively charged COA could be selectively bound. No protein sorption from the binary mixture was observed at pH = 4.0 for both PEM, when COA, LYZ, and the outermost PEI layer of PEM-5 were positively charged or the outermost PAC layer of PEM-6 was neutral. Furthermore, from factor analysis of the spectral data the higher selectivity was found for PEM-5 compared to PEM-6. Increasing the ionic strength revealed a drastic decrease in the selectivity of both PEM. Evidence was found that the proteins were predominantly bound at the surface and to a minor extent in the bulk phase of PEM. These results suggest possible working regimes and application fields of PEI/PAC multilayer assemblies related to the preparative separation of binary and multicomponent protein mixtures (biofluids, food) as well as to the design of selective protein-resistant surfaces.
Müller
M
M
Leibniz-Institute of Polymer Research Dresden e.V. (IPF), Hohe Strasse 6, D-01069 Dresden, Germany. mamuller@ipfdd.de
Kessler
B
B
Houbenov
N
N
Bohatá
K
K
Pientka
Z
Z
Brynda
E
E
D000180:Acrylic Resins / Q000737:chemistry*; D000327:Adsorption; D020650:Combinatorial Chemistry Techniques / Q000295:instrumentation* / Q000379:methods; D003208:Concanavalin A / Q000737:chemistry*; D004573:Electrolytes / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D008567:Membranes, Artificial; D009113:Muramidase / Q000737:chemistry*; D011094:Polyethyleneimine / Q000737:chemistry*; D012680:Sensitivity and Specificity; D017550:Spectroscopy, Fourier Transform Infrared / Q000295:instrumentation / Q000379:methods; D055672:Static Electricity; D013499:Surface Properties
D000180:Acrylic Resins; D004573:Electrolytes; D008567:Membranes, Artificial; D003208:Concanavalin A; C006903:carbopol 940; D011094:Polyethyleneimine; D009113:Muramidase