Langmuir monolayer approaches to protein recognition through molecular imprinting.
Surface plasmon resonance (SPR) spectroscopy and atomic force microscopy (AFM) have been employed to investigate ferritin adsorption to binary surfactant monolayers of cationic dioctadecyldimethylammonium bromide (DOMA) and non-ionic methyl stearate (SME). Surfactant molar ratios, miscibility, and lateral mobility were controlled to define the number, size, and distribution of "binding sites" for ferritin, which under the low ionic strength conditions investigated, adsorbed to the monolayers predominantly through electrostatic interactions. Successive adsorption/desorption cycles revealed that fluid monolayers, capable of laterally restructuring during the initial protein adsorption event, bound up to 60% more ferritin (dependent on SME:DOMA ratios) as compared to monolayers that were immobilized on a hydrophobic support during this first adsorption step. The enhanced binding of ferritin to fluid monolayers was accentuated in films having non-ionic SME as the principal component. These findings support the premise that the surfactants reorganize to form favorable interactions with an adsorbing protein, leading to protein specific charge patterns, or templates, in the films. Template assessment, however, was complicated by the presence of an irreversibly bound protein fraction, which AFM revealed to be locally ordered protein clusters.
Du
Xuezhong
X
Department of Bioengineering, University of Utah, Salt Lake City, UT 84112, USA.
Hlady
Vladimir
V
Britt
David
D
D000327:Adsorption; D015374:Biosensing Techniques / Q000379:methods*; D020099:Coated Materials, Biocompatible / Q000737:chemistry*; D005293:Ferritins / Q000032:analysis / Q000737:chemistry* / Q000648:ultrastructure*; D008422:Materials Testing; D018625:Microscopy, Atomic Force; D011485:Protein Binding; D025941:Protein Interaction Mapping / Q000379:methods*; D013228:Stearates / Q000032:analysis / Q000737:chemistry*; D020349:Surface Plasmon Resonance; D013499:Surface Properties; D013501:Surface-Active Agents / Q000032:analysis / Q000737:chemistry*
D020099:Coated Materials, Biocompatible; D013228:Stearates; D013501:Surface-Active Agents; D005293:Ferritins