title: Benefits and limitations of porous substrates as biosensors for protein adsorption.
abstract: Porous substrates have gained widespread interest for biosensor applications based on molecular recognition. Thus, there is a great demand to systematically investigate the parameters that limit the transport of molecules toward and within the porous matrix as a function of pore geometry. Finite element simulations (FES) and time-resolved optical waveguide spectroscopy (OWS) experiments were used to systematically study the transport of molecules and their binding on the inner surface of a porous material. OWS allowed us to measure the kinetics of protein adsorption within porous anodic aluminum oxide membranes composed of parallel-aligned, cylindrical pores with pore radii of 10-40 nm and pore depths of 0.8-9.6 μm. FES showed that protein adsorption on the inner surface of a porous matrix is almost exclusively governed by the flux into the pores. The pore-interior surface nearly acts as a perfect sink for the macromolecules. Neither diffusion within the pores nor adsorption on the surface are rate limiting steps, except for very low rate constants of adsorption. While adsorption on the pore walls is mainly governed by the stationary flux into the pores, desorption from the inner pore walls involves the rate constants of desorption and adsorption, essentially representing the protein-surface interaction potential. FES captured the essential features of the OWS experiments such as the initial linear slopes of the adsorption kinetics, which are inversely proportional to the pore depth and linearly proportional to protein concentration. We show that protein adsorption kinetics allows for an accurate determination of protein concentration, while desorption kinetics could be used to capture the interaction potential of the macromolecules with the pore walls.
authors.affiliation: Institute of Organic and Biomolecular Chemistry, Göttingen, Germany.
authors.forename: Thomas D
authors.initials: TD
authors.lastname: Lazzara
authors.forename: Ingo
authors.initials: I
authors.lastname: Mey
authors.forename: Claudia
authors.initials: C
authors.lastname: Steinem
authors.forename: Andreas
authors.initials: A
authors.lastname: Janshoff
mesh_terms: D000327:Adsorption; D000537:Aluminum Oxide / Q000737:chemistry* / Q000378:metabolism; D001360:Avidin / Q000737:chemistry* / Q000378:metabolism; D015374:Biosensing Techniques / Q000295:instrumentation*; D007700:Kinetics; D008956:Models, Chemical; D016062:Porosity; D011485:Protein Binding; D013499:Surface Properties
chemical_list: D001360:Avidin; D000537:Aluminum Oxide