Stable adhesion of phospholipid vesicles to modified gold surfaces.
Phospholipid vesicles are well-studied biomembrane mimics that are of increasing interest in drug delivery, immunoassays, and sensor chips. In a number of biosensor applications it is desirable to be able to adhere vesicles to a surface in a manner which does not result in their rupture or fusion. Such behavior should, in principle, be achievable by controlling the vesicle-surface and vesicle-vesicle interactions. We have varied vesicle composition and charge (phosphatidylcholine, phosphatidylcholine-phosphatidic acid 18 mol%) and solution ionic strength, to study the adhesion of fluorescent vesicles to glass, gold, and gold modified with chemisorbed acetyl-cysteine. The extent of chemisorption was characterized with angle-resolved X-ray photoelectron spectroscopy (ARXPS), and vesicle integrity and behavior was studied using entrapped and lipophilic fluorescent markers, together and in separate measurements. Vesicle fusion (by energy transfer), adhesion of intact vesicles (with entrapped calcein) and diffusion coefficients (by photobleaching recovery) were monitored using confocal fluorescence microscopy. Acetyl-cysteine modified gold surfaces were shown to be appropriate substrates for adhesion of intact vesicles. Finally, as a 'proof of principle' for fluorescence amplification, release of a self-quenching entrapped reporter dye (calcein) by the detergent Triton X-100 was followed in real time.
Sofou
Stavroula
S
Department of Chemical Engineering and Applied Chemistry, Columbia University, 500 West 120th Street, # 4721, New York, NY 10027, USA.
Thomas
James L
JL
D000327:Adsorption; D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods*; D020099:Coated Materials, Biocompatible / Q000138:chemical synthesis* / Q000737:chemistry; D004566:Electrodes; D005240:Feasibility Studies; D005452:Fluoresceins / Q000032:analysis; D006046:Gold / Q000737:chemistry*; D008081:Liposomes / Q000138:chemical synthesis / Q000737:chemistry*; D008561:Membrane Fusion; D010743:Phospholipids / Q000737:chemistry*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence / Q000295:instrumentation / Q000379:methods*
D020099:Coated Materials, Biocompatible; D005452:Fluoresceins; D008081:Liposomes; D010743:Phospholipids; D006046:Gold; C007740:fluorexon