Ultrafiltration membrane surfaces with grafted polymer 'tentacles': preparation, characterization and application for covalent protein binding.

Abstract

Ultrafiltration (UF) membranes from polysulfone (PSf) were functionalized by heterogeneous photo-initiated graft copolymerization of acrylic acid (AA). With radiation susceptible PSf, only proper selection of the UV energy (lambda > 350 nm; for selective excitation of the photoinitiator) yielded membranes with preserved UF barrier layer. Possibilities for adjusting structure and morphology of the graft polymer (g-PAA) layer by variation of functionalization parameters such as AA concentration and UV irradiation time were investigated. Very long grafted chains (Mw > 10(5) g mol(-1)) at varied grafting density (GD = 0.01 ... 1.2 nmol cm(-2), relative to the outer surface area) were obtained. Partial penetration of the UF barrier layer by g-PAA was verified. Covalent immobilization of bovine serum albumin (BSA), gamma-globulin (gamma-Gl) and alkaline phosphatase (APh) was achieved by coupling with a water soluble carbodiimide. Bound BSA and gamma-Gl amounts were up to gamma = 10 microg cm(-2), for membranes accessible only from the outer surface thus not using the entire pore volume. Locally addressed covalent protein immobilization after photo-patterning the PSf surface could be visualized with a fluorescent FITC-BSA conjugate. A strong salt effect onto immobilized APh activity (increase with NaCl concentration) was observed, indicating internal transport/accessibility limitations in the g-PAA layer. Correlations between PAA structure (Mw, GD) and accessibility (from BSA or gamma-G1 binding and APh activity) could be established. The 'tentacle' g-PAA functionalized PSf UF membranes having preserved UF barrier and, e.g., with surface-bound receptors will find application in cell cultures under diffusion or perfusion conditions.