Separation of Liposomes from Plasma Components Using Fast Protein Liquid Chromatography

Choice, Edward; Ayyobi, Amir F; Pritchard, P H

doi:10.1006/abio.1999.4049

Cited by 9 publications

(5 citation statements)

References 31 publications

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“…The elution profile of blank human plasma agrees well with previously published results and is shown in Fig. .…”

Section: Resultssupporting

confidence: 91%

Liposomes as solubilizers for lipophilic parenteral drugs: Transfer of drug and lipid marker to plasma proteins

Kaeß¹,

Fahr²

2014

Euro J Lipid Sci & Tech

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The effect of plasma on the retention of lipophilic drugs by liposomes as affected by the liposome fluidity was investigated. Liposomes consisted of eggPC/eggPG (9:1 w/w) or DPPC/DPPG (9:1 w/w) with 10 wt% incorporated 14C‐labeled Temoporfin (mTHPC) and trace amounts of 3H‐labeled cholesteryloleylether (COE) as lipid marker. Liposomal formulations were incubated at different concentrations in 50% human plasma for 48 h and the mixtures fractionated using gel permeation chromatography, separating liposomes and (lipo)proteins (albumin, HDL, LDL) from each other. The DPPC/DPPG liposomes retained 75–90% of the COE label at 48 h, depending on the liposome concentration, while the EPC/EPG liposomes retained only 5% of this label. By contrast, for the case of the mTHPC label, the EPC/EPG liposomes retained 24% at 48 h. whereas the DPPC/DPPG liposomes retained only 15%. In all cases, HDL was the predominant acceptor of both drug and COE label, closely followed by LDL. Albumin participated only marginally in the transfer process. It can be concluded, that EPC/EPG liposomes can retain lipophilic drugs better than DPPC/DPPG liposomes. Given a lifetime of 8 h for liposomes in vivo, 40% of the drug stays in the liposomes, whereas only 25% is retained for DPPC/DPPG liposomes at sufficient liposome concentration. Transfer of drug from liposomal formulations to human plasma was measured after separation of plasma components by GPC. Incubation of mTHPC loaded liposomes with human plasma results in a transfer of the drug and phospholipids mostly to HDL, followed by LDL, whereas albumin is only marginally involved in the transfer process. In comparison to a fluid liposomal formulation composed of eggPC/eggPG (9:1 w/w), rigid DPPC/DPPG (9:1 w/w) liposomes show less interactions with plasma proteins but a faster drug release.

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“…The elution profile of blank human plasma agrees well with previously published results and is shown in Fig. .…”

Section: Resultssupporting

confidence: 91%

Liposomes as solubilizers for lipophilic parenteral drugs: Transfer of drug and lipid marker to plasma proteins

Kaeß¹,

Fahr²

2014

Euro J Lipid Sci & Tech

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“…Fast Protein Liquid Chromatography (FPLC) was performed at 4°C using a Bio-Logic Pathfinder FPLC system interfaced with an automatic fraction collector (Bio-Rad, Hercules, CA) 26. See Supplemental Methods.…”

Section: Mterial and Methodsmentioning

confidence: 99%

Vascular-Directed Tissue Factor Pathway Inhibitor Overexpression Regulates Plasma Cholesterol and Reduces Atherosclerotic Plaque Development

Pan

White

Witt

et al. 2009

Circulation Research

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Rationale: Tissue factor pathway inhibitor (TFPI) is a potent regulator of the tissue factor pathway and is found in plasma in association with lipoproteins. Objective: To determine the role of TFPI in the development of atherosclerosis, we bred mice which overexpress TFPI into the apolipoprotein E-deficient (apoE ؊/؊ ) background. Methods and Results: On a high-fat diet, smooth muscle 22␣ (SM22␣)-TFPI/apoE ؊/؊ mice were shown to have less aortic plaque burden compared to apoE ؊/؊ mice. Unexpectedly, SM22␣-TFPI/apoE ؊/؊ had lower plasma cholesterol levels compared to apoE ؊/؊ mice. Furthermore, SM22␣-TFPI mice fed a high-fat diet had lower cholesterol levels than did wild-type mice. Because TFPI is associated with lipoproteins and its carboxyl terminus (TFPIct) has been shown to be a ligand for the very-low-density lipoprotein (VLDL) receptor, we hypothesized that TFPI overexpression may regulate lipoprotein distribution. We quantified VLDL binding and uptake in vitro in mouse aortic smooth muscle cells from SM22␣-TFPI and wild-type mice. Mouse aortic smooth muscle cells from SM22␣-TFPI mice demonstrated higher VLDL binding and internalization compared to those from wild-type mice. Because SM22␣-TFPI mice have increased circulating levels of TFPI antigen, we examined whether TFPIct may act to alter lipoprotein distribution. In vitro, TFPIct increased VLDL binding, uptake, and degradation in murine embryonic fibroblasts. Furthermore, this effect was blocked by heparinase treatment. In vivo, systemic administration of TFPIct reduced plasma cholesterol levels in apoE ؊/؊ mice. Conclusions: These studies suggest that overexpression of TFPI lowers plasma cholesterol through the interaction of its carboxyl terminus with lipoproteins and heparan sulfate proteoglycans. (Circ Res. 2009;105:713-720.)Key Words: atherosclerosis Ⅲ coagulation Ⅲ murine models Ⅲ tissue factor Ⅲ lipoproteins T issue factor (TF) pathway inhibitor (TFPI), a Kunitz-type serine protease inhibitor, is an endogenous inhibitor of TF-mediated coagulation. TFPI suppresses factor Xa generation by binding via its Kunitz 1 domain to the TF:FVIIa catalytic complex and via its Kunitz 2 domain to factor Xa. 1 The formation of a quaternary TF:VIIa:TFPI:Xa complex dampens ongoing FXa generation. Additionally, TFPI also has been shown to regulate coagulation by inducing the internalization and degradation of TF:VIIa complex on cell surfaces. [2][3][4] TFPI is expressed in many cells relevant to atherosclerosis including platelets, endothelial cells, vascular smooth muscle cells, and monocyte/macrophages. [5][6][7][8][9] In human carotid plaques, the level of TFPI expression is inversely associated with TF activity, suggesting a local regulatory role. 10 In plasma, TFPI exists in small quantities (Ͻ5%) as a free full-length protein but is predominantly associated with lipoproteins. 11-13 Lipoprotein-associated TFPI has been shown to have less anticoagulant activity than free TFPI. 13 TFPI is cleared from the plasma by binding the low-density lipoprotein recepto...

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“…Liposomes are typically separated from other biological compounds with techniques varying from one team to another, such as ultracentrifugation coupled with membrane ultrafiltration 133 . Another example is the chromatographic separation, specifically fast protein liquid chromatography, which enables the separation of liposomes from lipoproteins and plasma components 134 . Several studies analyzing the formation of the in vivo PC led to the same conclusion as in vitro.…”

Section: Parameters Influencing the Pc In Vivomentioning

confidence: 99%

“… 133 Another example is the chromatographic separation, specifically fast protein liquid chromatography, which enables the separation of liposomes from lipoproteins and plasma components. 134 …”

Section: Studying Pc In Vivo : the Way To Improve ...mentioning

confidence: 99%

In vivo protein corona on nanoparticles: does the control of all material parameters orient the biological behavior?

et al. 2021

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Separation of Liposomes from Plasma Components Using Fast Protein Liquid Chromatography

Cited by 9 publications

References 31 publications

Liposomes as solubilizers for lipophilic parenteral drugs: Transfer of drug and lipid marker to plasma proteins

Liposomes as solubilizers for lipophilic parenteral drugs: Transfer of drug and lipid marker to plasma proteins

Vascular-Directed Tissue Factor Pathway Inhibitor Overexpression Regulates Plasma Cholesterol and Reduces Atherosclerotic Plaque Development

In vivo protein corona on nanoparticles: does the control of all material parameters orient the biological behavior?

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