Dspe peg2000

Manufactured by Avanti Polar Lipids

Sourced in United States, Germany

DSPE-PEG2000 is a lipid conjugate compound composed of distearoylphosphatidylethanolamine (DSPE) and polyethylene glycol (PEG) with an average molecular weight of 2000 Da. It is a widely used material in the development of liposomal drug delivery systems and other biomedical applications.

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Close spelling variants of this product

1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000] (DSPE-PEG2000) (32 citations) 1,2-Distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-distearoyl-sn-glycero-3-phospho-ethanol amine-N-[methoxy(polyethylene glycol)-2000] (ammonium salt) (DSPE-PEG2000), (2 citations) DSPE-PEG2000-MAL (19 citations) 1,2-distearoryl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethyleneglycol-2000)] ammonium salt (DSPE-PEG2000) (2 citations) DSPE-Bio-PEG2000 (3 citations) 1,2-Distearoyl-sn-glycero-3-phosphoethanolamine-N-carboxy (polyethylene glycol) 2000 (DSPE–PEG2000–COOH) (3 citations) 1,2-dipalmitoyl-sn-glycero-3-phosphocho-line (DPPC), 1,2-distearoyl-sn-glycero-3-phosphoethanola-mine-N-[amino(polyethyleneglycol)-2000] (DSPE-PEG2000-Amine), (2 citations) H2N-PEG2000-DSPE (2 citations) 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-(polyethylene glycol 2000) (DSPE-PEG2000) (4 citations) 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[maleimide(polyethylene glycol)-2000] (DSPE-PEG2000-MAL), (5 citations)

149 protocols using dspe peg2000

Synthesis of DDFP-Encapsulated Lipid Droplets

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Dodecafluoropentane (DDFP) from Synquest Labs (Alachua, FL, USA) was used as the core of PC droplets. DDFP has a boiling point of 29 °C at atmospheric pressure. The lipid shell encapsulating the PC droplet was fabricated using 1,2-ditearoyl-sn-glycero-3-phosphocholine (DSPC) and 1,2-dioctadecanoyl-sn-glycero-3-phosphoethanolamine-polyethylene glycol-2000 (DSPE-PEG2000), which were purchased from Avanti Polar Lipids (Alabaster, AL, USA).
DSPC and DSPE-PEG2000 were dissolved in chloroform at molar ratios of 9:1 or 7:3 in a 2 mL clear vial. The vial was placed in a vacuum chamber for 12 h to completely remove the chloroform. Subsequently, a thin white lipid film formed at the bottom of the vial was hydrated with 1.8 mL of distilled water and sonicated using a bath sonicator (Powersonic 505, Hwashin Instrument, Seoul, Republic of Korea) at 56 °C. After allowing the vial to rest with the lipid solution at room temperature, liquid DDFP was added to the vial to achieve a volume fraction of 10% [16 (link)]. Finally, DDFP droplets were created by agitating the vials for 45 s in a Vialmix (Lantheus Holdings, Billerica, MA, USA).

Jeong S.Y., Seo H.B., Seo M.H., Cho J.W., Kwon S., Son G, & Lee S.Y. (2024). Repeatable Acoustic Vaporization of Coated Perfluorocarbon Bubbles for Micro-Actuation Inspired by Polypodium aureum. Biomimetics, 9(2), 106.

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Fluorine-rich Nanoparticle Synthesis and Functionalization

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Nanoparticles with high fluorine content were prepared by emulsifying Perfluoro-15-crown-5-ether (PFCE, Fluorochem, Derbyshire, UK) via direct sonication, using a ultrasonic homogenizer (Hielscher Ultrasonic GmbH, Teltow, Berlin, Germany). PFCE was emulsified in Pluronic F-68 (Sigma-Aldrich, Germany) for 10 minutes on ice (1.2 M end concentration) to generate a basic formulation: Basic ¹⁹F nanoparticles (NP). To enrich the basic particles with DPPE (1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine, Figure 1A, Avanti Polar Lipid, Inc., Alabaster, AL) diluted Pluronic-basic nanoparticles were mixed with DPPE and further emulsified using the same conditions to obtain a final PFCE concentration of 400 mM and varying DPPE concentrations of 2.5–25 μM (DPPE ¹⁹F NP). The same procedure was used to prepare the Pluronic-Rhodamine-DPPE (DPPE-Rhodamine ¹⁹F NP) and nanoparticles enriched with POPE (1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine, Figure 1A, Avanti Polar Lipid, Inc., Alabaster, AL). DSPE-PEG2000—PFCE nanoparticles were prepared by first coating a vial with 10 mg of DSPE-PEG2000 (1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N[methoxy(polyethylene glycol) -2000]), Avanti Polar Lipid, Inc., Alabaster, AL) and then sonicating PFCE in the vial for 10 minutes on ice, to achieve a final concentration of 120 mM PFCE and 3.75 mM DSPE-PEG2000.

Waiczies S., Lepore S., Sydow K., Drechsler S., Ku M.C., Martin C., Lorenz D., Schütz I., Reimann H.M., Purfürst B., Dieringer M.A., Waiczies H., Dathe M., Pohlmann A, & Niendorf T. (2015). Anchoring Dipalmitoyl Phosphoethanolamine to Nanoparticles Boosts Cellular Uptake and Fluorine-19 Magnetic Resonance Signal. Scientific Reports, 5, 8427.

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Thermosensitive Liposomal Doxorubicin Preparation

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Thermosensitive Liposomal Doxorubicin (TSL-Dox) was prepared as described earlier [24 (link)]. Briefly 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), monostearoylphosphatidylcholine (MSPC), and 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-PEG2000 (DSPE-PEG2000) (Avanti Lipids, Alabaster, AL, USA) (DPPC:MSPC:DSPE-PEG2000 = 85.3:9.7:5.0 mol%) were dissolved in chloroform and dried under a stream of atmospheric air at room temperature for forming a thin film. The lipids were then hydrated with 300 mM citric acid buffer (pH 4.0) and extruded 5 times at 55 °C (Thermobarrel extruder; Northern Lipids, BC, Canada) through a 100 nm filter. Active loading of doxorubicin into the liposomes was carried out by pH gradient with phosphate buffered saline (PBS, pH 7.4) outside the liposomes. The release kinetics of doxorubicin from the TSL was measured between 37 and 45 °C by a millifluidic device, as described earlier [42 (link),43 ].
A second batch of TSL was prepared based on the same lipid composition as above, with the addition of 0.1 mol% of Rhodamine headgroup-labeled 1,2-Dipalmitoyl-sn-glycero-3-phosphoethanolamine (Rho-DPPE) to produce empty, fluorescence-labeled TSL. This second batch was used to quantify the removal of TSL by the filter.

Motamarry A., Wolfe A.M., Ramajayam K.K., Pattanaik S., Benton T., Peterson Y., Faridi P., Prakash P., Twombley K, & Haemmerich D. (2022). Extracorporeal Removal of Thermosensitive Liposomal Doxorubicin from Systemic Circulation after Tumor Delivery to Reduce Toxicities. Cancers, 14(5), 1322.

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Liposomal Nanocarrier Preparation for Therapeutic Delivery

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Liposomal nanocarriers were prepared with Dimyristoyl-sn-glycero-3-phosphocholine (DMPC) and pegylated distearoyl-phosphatidyl ethanolamine (DSPE-PEG-2000) (Avanti Lipids) as previously described (Hamurcu et al., 2016 (link)). Briefly, DMPC and DSPE-PEG2000 were mixed at the ratio of (10:1) and were mixed with small molecule inhibitors at a ratio of 10:1 (w/w) and lyophilized in the presence of excess tertiary butanol. After liposomal drugs were reconstituted in PBS, the agents were systemically administered to mice.

Yan G., Luna A., Wang H., Bozorgui B., Li X., Sanchez M., Dereli Z., Kahraman N., Kara G., Chen X., Zheng C., McGrail D., Sahni N., Lu Y., Babur O., Cokol M., Lim B., Ozpolat B., Sander C., Mills G.B, & Korkut A. (2022). BET inhibition induces vulnerability to MCL1 targeting through upregulation of fatty acid synthesis pathway in breast cancer. Cell reports, 40(11), 111304.

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Pegylated Liposomal Delivery of miRNA

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For in vivo delivery, miRNA was incorporated into pegylated liposomes composed of dimyristoyl-sn-glycero-3-phosphocholine (DMPC) and pegylated distearoyl-phosphatidylethanolamine (DSPE-PEG-2000) (Avanti Lipids). DMPC and DSPE-PEG2000 were mixed at a 10:1 ratio, and miRNA (control or miR-603 mimic, 8ug miR/mouse) was mixed at a 10:1 (w/w) ratio of lipids to oligonucleotides in the presence of excess tertiary butanol. Prior to in vivo administration, lyophilized lipid/miRNA complex was reconstituted in 0.9% saline. The liposomal suspension filtered and centrifuged. The liposomes trapped in the filter was reconstituted and used for injections.

Bayraktar R., Pichler M., Kanlikilicer P., Ivan C., Bayraktar E., Kahraman N., Aslan B., Oguztuzun S., Ulasli M., Arslan A., Calin G., Lopez-Berestein G, & Ozpolat B. (2016). MicroRNA 603 acts as a tumor suppressor and inhibits triple-negative breast cancer tumorigenesis by targeting elongation factor 2 kinase. Oncotarget, 8(7), 11641-11658.

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DMPC-DSPE-PEG2000 Liposomal siRNA Delivery

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In vivo siRNA delivery was achieved by incorporating siRNA into liposomes consisted of Dimyristoyl-sn-glycero-3-phosphocholine (DMPC) and pegylated distreroly-phosphotidyl ethanolamine (DSPE-PEG-2000) (Avanti Lipids). DMPC and DSPE-PEG2000 were mixed at the ratio of (10:1) and siRNA were mixed at a ratio of 10:1 lipid to oligos (control siRNAor FOXM1 siRNA) (w/w) in the presence of excess tertiary butanol. Prior to in vivo administration, the siRNA/lipid complex was reconstituted in saline and systemically administered (0.3 mg/kg or 8 μg/mouse) once a week in a volume of 100 μl.

Hamurcu Z., Ashour A., Kahraman N, & Ozpolat B. (2016). FOXM1 regulates expression of eukaryotic elongation factor 2 kinase and promotes proliferation, invasion and tumorgenesis of human triple negative breast cancer cells. Oncotarget, 7(13), 16619-16635.

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Preparation of Perfluorocarbon-Stabilized Nanoemulsions

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The lipids 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) and 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[amino(polyethylene glycol)-2000] (DSPE-PEG2000) were purchased from Avanti Lipids (Alabaster, AL, USA) and liquid perfluorocarbon (perfluoropentane) was obtained from Fluoromed (Round Rock, TX, USA). Lipids (190 mg) were dissolved in chloroform in a molar ratio of 94:6 DPPC:DSPE-PEG2000. The solvent was evaporated, and the dry lipid film was rehydrated with 10 ml phosphate-buffered saline. The lipid solution was sonicated for 2 minutes at 20% power using a Vibra-Cell sonicator probe (Sonics & Materials, Newtown, CT, USA) before adding liquid perfluoropentane (C₅F₁₂) at a volume ratio of 4%. The mixture was sonicated at 25% power in pulsed mode (10 s on, 50 s off) in an ice-water bath for a total sonication time of 60 s, followed by 16 passes through 200 nm polycarbonate membrane filters (Whatman, Piscatway, NJ, USA) using a LIPEX extruder (Northern Lipids, Burnaby, BC, Canada). The mean size and polydispersity of the droplets measured using a particle size analyzer (90Plus Particle Size Analyzer, Brookhaven Instruments, Holtsville, NY, USA) was 169 ± 22 nm and 0.17 ± 0.05, respectively. The resulting PSNE solutions were stored at 4 °C until use.

Kopechek J.A., Park E.J., Zhang Y.Z., Vykhodtseva N.I., McDannold N.J, & Porter T.M. (2014). Cavitation-enhanced MR-guided focused ultrasound ablation of rabbit tumors in vivo using phase shift nanoemulsions. Physics in medicine and biology, 59(13), 3465-3481.

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Liposomal Nanocarrier Preparation for Therapeutic Delivery

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Lipid-based Nanoparticle Formulation and Evaluation

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Soybean phosphatidylcholine (SPC), cholesterol, and DSPE-PEG2000 were purchased from Avanti lipids (Avanti Polar Lipids, Alabaster, AL, USA). Dimethyl sulfoxide (DMSO), copper chloride (CuCl₂), and 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Fetal bovine serum (FBS) and DMEM-F12 medium (access date 12 March 2021,Gibco, Grand Island, NY, USA) were purchased from Thermo Fisher Scientific (Waltham, MA, USA), respectively. DQ was synthesized in our laboratory based on a previous report [12 (link)].

Kang X., Wang Q., Wu S., Wang C., Annaji M., Huang C.H., Shen J., Chen P, & Babu R.J. (2023). Liposomal DQ in Combination with Copper Inhibits ARID1A Mutant Ovarian Cancer Growth. Biomolecules, 13(5), 744.

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Producing Targeted and Non-Targeted Microbubbles

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Both CXCR4-targeted (T140-MB) and non-targeted (NT-MB) MB were produced in-house. The NT-MB’s shell contained a mixture of DPPC, DPPA and DSPE-PEG₂₀₀₀-NH₂ (81.9:8.6:9.5 mole ratio, respectively) (Avanti Lipids, Alabaster, AL, USA) (S1 Table) and in the T140-MB, 4.8% of the DSPE-PEG₂₀₀₀-NH₂ was replaced by the modified lipid, DSPE-PEG₂₀₀₀-T140 (Fig 1A and S2 Table). These components were dissolved in CHCl₃ and aliquoted into 3 mL vials from which the solvent was dried using a N₂ flow and stored at -20⁰C. Before use, the vials were reconstituted with propylene glycol, phosphate-buffered saline (PBS) and glycerol (16:5:1) whilst stirring. After degassing for a minimum of 15 min, the vial was crimp-sealed and the headspace of the vial was gas-exchanged with 99.0% n-perfluorobutane (FluoroMed L.P., Round Rock, TX, USA). MB were formed via standard mechanical agitation techniques using a Vialmix shaker (Bristol-Myers-Squibb, NY, USA) (2x30 sec shaking). Fluorescent MB were developed by adding 2 μg/mL of Dil stain (1,1’-Dioctadecyl-3,3,3’,3’-Tetramethylindocarbocyanine perchlorate, DilC₁₈(3), Thermo Fisher Scientific, Carlsbad, CA USA).

Braga M., Leow C.H., Gil J.H., Teh J.H., Carroll L., Long N.J., Tang M.X, & Aboagye E.O. (2021). Investigating CXCR4 expression of tumor cells and the vascular compartment: A multimodal approach. PLoS ONE, 16(11), e0260186.

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