Immobilization of liposomes onto quartz crystal microbalance to detect interaction between liposomes and proteins.
To study the interaction between liposomes and proteins, intact liposomes were immobilized on a metal planar support by chemical binding and/or bioaffinity using a quartz crystal microbalance (QCM). A large decrease in the resonance frequency of quartz crystal was observed when the QCM, modified by a self-assembled monolayer (SAM) of carboxythiol, was added to liposome solutions. The stable chemical immobilization of intact liposomes onto SAM was judged according to the degree with which adsorbed mass depended on the prepared size of liposomes, as well as on the activation time of SAMs when amino-coupling was introduced, where the liposome coverage of electrodes was 69+/-8% in optimal conditions. When avidin-biotin binding was used on amino-coupling liposome layers, liposome immobilization finally reached 168% coverage of the electrode surface. Denatured protein was also successfully detected according to the change in the frequency of the liposome-immobilized QCM. The adsorbed mass of denatured carbonic anhydrase from bovine onto immobilized liposomes showed a characteristic peak at a concentration of guanidine hydrochloride that corresponded to a molten globule-like state of the protein, although the mass adsorbed onto deactivated SAM increased monotonously.
Morita
Seiichi
S
Department of Materials Science, Wakayama National College of Technology, Noshima 77, Nada-cho, Gobo, Wakayama 644-0023, Japan. morita@wakayama-nct.ac.jp
Nukui
Miho
M
Kuboi
Ryoichi
R
D000327:Adsorption; D000818:Animals; D001360:Avidin / Q000378:metabolism; D015374:Biosensing Techniques; D001710:Biotin / Q000378:metabolism; D002256:Carbonic Anhydrases / Q000378:metabolism; D002417:Cattle; D006046:Gold; D007103:Immobilization; D007700:Kinetics; D008051:Lipid Bilayers / Q000737:chemistry; D008081:Liposomes / Q000737:chemistry* / Q000378:metabolism; D011506:Proteins / Q000737:chemistry / Q000378:metabolism*; D011791:Quartz; D013499:Surface Properties; D013816:Thermodynamics
D008051:Lipid Bilayers; D008081:Liposomes; D011506:Proteins; D001360:Avidin; D011791:Quartz; D001710:Biotin; D006046:Gold; D002256:Carbonic Anhydrases