Formation of protein molecular imprints within Langmuir monolayers: a quartz crystal microbalance study.
Protein imprinting leading to enhanced rebinding of ferritin to ternary lipid monolayers is demonstrated using a quartz crystal microbalance. Monolayers consisting of cationic dioctadecyldimethylammonium bromide, non-ionic methyl stearate, and poly(ethylene glycol) bearing phospholipids were imprinted with ferritin at the air/water interface of a Langmuir-Blodgett trough and transferred hydrated to hydrophobic substrates for study. This immobilization was shown by fluorescence correlation spectroscopy to significantly hinder any further diffusion of lipids, while rebinding studies demonstrated up to a six-fold increase in ferritin adsorption to imprinted versus control monolayers. A diminished rebinding of ferritin to its imprint was observed through pH reduction to below the protein isoelectric point, demonstrating the electrostatic nature of the interaction. Rebinding to films where imprint pockets remained occupied by the template protein was also minimal. Studies with a smaller acidic protein revealed the importance of the steric influence of poly(ethylene glycol) in forming the protein binding pockets, as albumin-imprinted monolayers showed low binding of ferritin, while ferritin-imprinted monolayers readily accommodated albumin. The controllable structure-function relationship and limitations of this system are discussed with respect to the application of protein imprinting in sensor development as well as fundamental studies of proteins at dynamic interfaces.
Turner
Nicholas W
NW
Department of Bioengineering, University of Utah, Salt Lake City, UT 84112, USA.
Wright
Bryon E
BE
Hlady
Vladimir
V
Britt
David W
DW
D000327:Adsorption; D000418:Albumins / Q000737:chemistry*; D000818:Animals; D015374:Biosensing Techniques; D005293:Ferritins / Q000737:chemistry*; D006736:Horses; D057927:Hydrophobic and Hydrophilic Interactions; D008055:Lipids / Q000737:chemistry*; D008567:Membranes, Artificial; D018625:Microscopy, Atomic Force; D011485:Protein Binding; D011791:Quartz / Q000737:chemistry; D055672:Static Electricity
D000418:Albumins; D008055:Lipids; D008567:Membranes, Artificial; D011791:Quartz; D005293:Ferritins