Conditions for liposome adsorption and bilayer formation on BSA passivated solid supports

Silva-Lopez, Elsa; Edens, Lance E.; Barden, Adam O.; Keller, David L.; Brozik, James A.

doi:10.1016/j.chemphyslip.2014.06.002

Cited by 15 publications

(12 citation statements)

References 62 publications

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“…To overcome the limitation that hindered the application of lipoproteins, BSA, a versatile protein with great solubility which has shown to be biocompatible, extremely robust (stable in the pH range of 4-9) and readily accessible, 12 was chosen as an alternative for apolipoprotein to construct novel biomimetic recostituted lipoproteins (rLips). 13,14 In addition, as a hydrophilic endogenous substance, BSA could prolong systemic circulation in vivo through minimizing the association with serum proteins and clearance of reticuloendothelial system (RES). 15,16 Besides, BSA may target tumor cells through the GP60 receptor and SPARC-mediated endocytosis.…”

Section: Introductionmentioning

confidence: 99%

<p>Lipoprotein-Inspired Nanocarrier Composed of Folic Acid-Modified Protein and Lipids: Preparation and Evaluation of Tumor-Targeting Effect</p>

Han

et al. 2020

IJN

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Background: Reconstituted lipoproteins (rLips) based on endogenous lipid nanostructures has been increasingly regarded as an excellent and promising antitumor drug delivery. However, some problems relating to the main component, apolipoprotein, for instance, rare source, unaffordable price, and low specificity of relevant receptor expression, become chief obstacles to its broad development and application. Purpose: The primary aim of this study is to develop biomimetic rLips by utilizing folic acid (FA)-modified bovine serum albumin (BSA) as a replacement for apolipoprotein and demonstrate its tumor targeting and antitumor efficacy. Methods: The amino groups of BSA were covalently conjugated with FA through the amide reaction. PTX-loaded nanostructured lipid carrier (termed as P-NLC) consisting of phospholipid, cholesteryl ester, triglyceride and cholesterol was prepared by the emulsification-evaporation method and utilized as the lipid core. FA-modified BSA (FA-BSA) was characterized for the protein substitute degree and attached with NLC by incubation-insert method to form the lipoprotein-mimic nanocomplex (termed as PFB-rLips). The morphology of nanoparticles was observed under transmission electron microscopy (TEM), and the particle size and zeta potential were determined using dynamic light scattering. In vitro release behavior of PTX from PFB-rLips was investigated with the dialysis method. Hemolysis tests were conducted to evaluate the biosecurity of PFB-rLips. Cell uptake and cytotoxicity assays were performed on human hepatocytes (LO2) and human hepatoma cells (HepG2). Tumor targeting was assessed using in vivo imaging system in H22 tumor-bearing mice model. Antitumor efficacy in vivo was investigated and compared between Taxol ® (paclitaxel) formulation and PTX-incorporated nanoparticles in the same tumor model. Results: A fixed molar ratio 50:1 of FA to BSAwas chosen as the optimal input ratio based on the balance between appropriate degree of protein substitution and amphiphilicity of FA-BSA. The morphology of FB-rLips exhibited as a homogeneous spherical structure featured by lipid cores surrounded with a cloudy protein shell observed under TEM. The particle size, zeta potential and encapsulation efficiency were 174.6±3.2 nm, −17.26±0.9 mVand 82.2±2.4%, respectively. In vitro release behavior of PTX from PFB-rLips was slow and sustained. The uptake of FB-rLips was much higher in HepG2 cells than in LO2 cells. Furthermore, the uptake of FB-rLips was significantly higher than that of rLips without FA involved (termed as B-rLips) and NLC in HepG2 cells. Hemolysis and cytotoxicity assays showed good biocompatibility of FB-rLips. The internalization mechanism of FB-rLips mainly depended on clathrin-mediated and caveolinmediated endocytosis coupling with energy consumption, and FA receptors expressed on tumor cells played a critical role in cellular uptake process. CCK-8 studies demonstrated that PFB-rLips

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Section: Introductionmentioning

confidence: 99%

<p>Lipoprotein-Inspired Nanocarrier Composed of Folic Acid-Modified Protein and Lipids: Preparation and Evaluation of Tumor-Targeting Effect</p>

Han

et al. 2020

IJN

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“…The solution outside the liposome (External Buffer, EB) was prepared by mixing PBS (1X), BSA solution (100 mg mL −1 ) and 20 % glucose in water in PBS. This external buffer mainly promotes the bilayer liposome preparation in reverse emulsion technique . Osmolarity of EB and IB was measured by Osmometer in our lab.…”

Section: Methodsmentioning

confidence: 99%

“…This external buffer mainly promotes the bilayer liposome preparation in reverse emulsion technique. [44] Osmolarity of EB and IB was measured by Osmometer in our lab. Osmolarity of IB was kept approximately 5-10 mOsm higher than that of the EB to maintain the shape of the liposome.…”

Section: Methodsmentioning

confidence: 99%

Probing Deviation of Adhered Membrane Dynamics between Reconstituted Liposome and Cellular System

Mondal

Chowdhury

Nandi

et al. 2019

Chemistry An Asian Journal

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The dynamics of cell-cell adhesion are complicated due to complexities in cellular interactions and intra-membrane interactions. In the present work, we have reconstituted al iposome-based model system to mimic the cell-cell adhesion process. Our model liposome system consists of one fluorescein-taggeda nd one TRITC (tetramethyl-rhodamine isothiocyanate)-tagged liposome, adhered through biotinneutravidin interaction. We monitored the adhesion process in liposomes using Fçrster Resonance EnergyT ransfer (FRET) between fluorescein( donor) and TRITC (acceptor). Occurrence of FRET is confirmed by the decrease in donor lifetime as well as distinct rise time of the acceptorf luorescence. Interestingly,t he acceptor'se mission exhibits fluctuations in the range of % 3 AE 1s.T his may be attributed to structural oscillationsa ssociated in two adhered liposomesa rising from the flexible nature of biotin-neutravidin interaction. We have compared the dynamics in ac ell-mimicking liposome system with that in an in vitro live cell system.I nt he adhered live cell system, we used CPM (7-diethylamino-3-(4maleimido-phenyl)-4-methylcoumarin, donor) andn ile red (acceptor), whicha re known to stain the membrane of CHO (Chinese Hamster Ovary) cells. The dynamicso ft he adhered membranes of two live CHO cells were observed through FRET between CPM and nile red. The acceptorf luorescence intensity exhibits an oscillation in the time-scaleo f% 1 AE 0.75 s, which is faster comparedt ot he reconstituted liposome system,i ndicating the contributions and involvement of multiple dynamic protein complexes around the cell membrane. This study offers simple reconstituted model systems to understand the complex membrane dynamics using aF RET-based physical chemistry approach.[a] Dr.

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“…Under these conditions, liposomes fuse to the surface, forming a uniform supported lipid bilayer. 46 As a precaution to minimize non-specific binding, BSA was then added to samples at a concentration of 0.5 mg/mL and the samples were vortexed on low speed at room temperature for 45 minutes. Beads were then washed 4–5X to remove unbound liposomes by cycles of centrifugation at 10,000 rpm for 45 seconds, followed by removing the supernatant, always leaving at least 50 µL in the tube to keep the lipid coated beads hydrated.…”

Section: Methodsmentioning

confidence: 99%

Multiplexed Lipid Bilayers on Silica Microspheres for Analytical Screening Applications

et al. 2017

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Most druggable targets are membrane components, including membrane proteins and soluble proteins that interact with ligands or receptors embedded in membranes. Current target-based screening and intermolecular interaction assays generally do not include the lipid membrane environment in presenting these targets, possibly altering their native structure and leading to misleading or incorrect results. To address this issue, an ideal assay involving membrane components would (1) mimic the natural membrane environment, (2) be amenable to high-throughput implementation, and (3) be easily multiplexed. In a step toward developing such an ideal target-based analytical assay for membrane components, we present fluorescently indexed multiplexed biomimetic membrane assays amenable to high-throughput flow cytometric detection. We build fluorescently multiplexed biomimetic membrane assays by using varying amounts of a fluorescently labeled lipid, NBD-DOPE [1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-N-(7-nitro-2-1,3-benzoxadiazol-4-yl)], incorporated into a phospholipid membrane bilayer supported on 3 μm silica microspheres. Using flow cytometry, we demonstrate this multiplexed approach by measuring specific affinity of two well-characterized systems, the fluorescently labeled soluble proteins cholera toxin B subunit-Alexa 647 and streptavidin-PE/Cy5, to membranes containing different amounts of ligand targets of these proteins, GM1 and biotin-DOPE, respectively. This work will enable future efforts in developing highly efficient biomimetic assays for interaction analysis and drug screening involving membrane components.

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Conditions for liposome adsorption and bilayer formation on BSA passivated solid supports

Cited by 15 publications

References 62 publications

<p>Lipoprotein-Inspired Nanocarrier Composed of Folic Acid-Modified Protein and Lipids: Preparation and Evaluation of Tumor-Targeting Effect</p>

<p>Lipoprotein-Inspired Nanocarrier Composed of Folic Acid-Modified Protein and Lipids: Preparation and Evaluation of Tumor-Targeting Effect</p>

Probing Deviation of Adhered Membrane Dynamics between Reconstituted Liposome and Cellular System

Multiplexed Lipid Bilayers on Silica Microspheres for Analytical Screening Applications

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