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Dspe peg 2000 amine

Manufactured by Avanti Polar Lipids
Sourced in United States

DSPE-PEG-2000-Amine is a lipid-based compound used in various laboratory applications. It consists of 1,2-distearoyl-sn-glycero-3-phosphoethanolamine (DSPE) covalently linked to polyethylene glycol (PEG) of molecular weight 2000 and an amine functional group. This product is commonly used as a surfactant, emulsifier, or for the preparation of liposomes and other lipid-based nanoparticles.

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10 protocols using dspe peg 2000 amine

1

Synthesis of Cationic Lipid Nanoparticles

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A certain mass of lipid [5 mg DPPC, 2 mg DSPE-PEG (2000)-Amine and 2 mg DC-cholesterol(Avanti Polar Lipids, Inc, Alabaster, AL, USA)] were mixed together and dissolved into 10 mL trichloromethane(CHCl3) (Fisher Scientific, Waltham, MA, USA). The solution was moved to the rotary evaporator (Yarong Inc, Shanghai, China) at 50 °C to remove the organic solvent and form a thin lipid film. One hour later, the resulting thin lipid films was hydrated in 2 mL Double steaming water forming a translucent opalescent suspension. Then 100 µl of Perfluorohexane (PFH) was dropwise into suspension after which the suspension was emulsified in an ice bath using a sonicator (Sonics &Materials Inc., Newtown, CT, USA) with power of 130 W for 5 min (5 s on and 5 s off), then the prepared cationic liquid nanoparticles (CL-NPs) were harvested. The nanoemulsions were centrifuged at 6000 rpm for 5 min and then washed in Double steaming water to wipe off disasociated lipids and PFH. The centrifugation and washing process were repeated three times. Finally, the prepared CL-NPs were stored at 4 °C for later use. To prepare fluorescent nanodroplets, DiI (1 mg) or DiR (1 mg) fluorescent dye were added to the lipid solution and aluminum foil papers was used to prevent light exposure.
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2

Synthesis and Characterization of ART-1-Lipopeptide

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IL-27 was purchased from PeproTech, USA. DOPC (1, 2-Dioleoyl-sn-glycero-3-phosphocholine), DOPE (1, 2-Dioleoyl-sn-glycero-3-phosphoethanolamine), cholesterol and FITC (fluorescein isothiocyanate) were obtained from Sigma-Aldrich, USA. DSPE-PEG (2000) amine (1, 2-distearoyl-sn-glycero-3-phosphoethanolamine-N- [amino (polyethylene glycol)-2000] (ammonium salt)) was purchased from Avanti Polar Lipids, USA. Cyanine 7 azide (Cy7) was purchased from Lumiprobe, USA. ART-1-lipopeptide (CRNADKFPC-NH-C18H37) was customsynthesized by GenScript/Lifetein, USA. The structure of ART-1-lipopeptide is shown in Fig. S1. The peptide in both the lipopeptide and liposomal formulation used in this study was found to be in cyclic form (C-C disulfide 1–9) as assessed by LC- MS spectra of ART-1-lipopeptide: m/z calculated 1301.74 for CRNADKFPC-NH-C18H37, found 651.88 ([M+H]+/2) and 434.92 ([M+H]+/3) (protonated molecular ions of the formula (M+H)+ in positive ionization mode).
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3

Lipid-based Nanoparticle Fabrication

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Lactel® 50:50 PLGA was purchased from Durect Corporation (Cupertino, CA). Fetal bovine serum (FBS), granulocyte macrophage-colony stimulating factor (GM-CSF) recombinant mouse protein, alpha minimum essential medium, trypsin/EDTA, Alexa Fluor® 647 hydrazide, and tris(triethylammonium) salt were purchased from Life Technologies Corporation (Grand Island, NY). Lipids, including 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[amino(polyethylene glycol)-2000] (ammonium salt) ((DSPE-PEG(2000)) amine), cholesterol, and 1,2-diphytanoyl-sn-glycero-3-phosphoethanolamine-N-(7-nitro-2-1,3-benzoxadiazol-4-yl) (ammonium salt) (NBD PE) were purchased from Avanti Polar Lipids, Inc. (Alabaster, AL). Poly (vinyl alcohol) (PVA, MW 89,000–98,000), dichloromethane (DCM), rhodamine B (Rhod B), and BSA were purchased from Sigma-Aldrich Inc. (Saint Louis, MO). 1-Ethyl-3-[3-dimethylaminopropyl] carbodiimide hydrochloride (EDC) was purchased from Thermo Fisher Scientific Inc. (Rockford, IL). JAWSII (ATCC® CRL-11904™) immature dendritic cells were purchased from ATCC (Manassas, VA). All other chemicals were of analytical grade.
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4

Lipid-Based Nanoparticle Functionalization

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1,2-Distearoyl-sn-glycerol-3-phosphocholine (DSPC), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[biotinyl(polyethylene glycol)-2000] (ammonium salt) (DSPE-PEG-2000-Biotin), and 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[amino(polyethylene glycol)-2000] (ammonium salt) (DSPE-PEG-2000-Amine) were purchased from Avanti Polar Lipids (Alabaster, AL). IL-2, IL-6, and TNF-α ELISA kits were purchased from BioLegend (San Diego, CA). Cryopreserved human PBMCs were purchased from Cellular Technology Ltd (Cleveland, OH). FITC-avidin was purchased from Thermo Fisher Scientific (Waltham, MA). Dulbecco's phosphate-buffered saline (DPBS), chloroform, LPS, and MPA were purchased from Sigma Aldrich (St. Louis, MO). The anti-HIF-1ɑ antibody was purchased from Santa Cruz Biotechnology, Inc. (Dallas, TX).
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5

Bifidobacterium longum Nanoparticle Formulation

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Bifidobacterium longum (ATCC 15707) was obtained from the Chongqing Key Laboratory of Biomedical Engineering. Man-Rogosa-Sharpe (MRS) broth was purchased from Qingdao Haibo Biotechnology Co. (Qingdao, China). 4,6-Diamidino-2-phenylindole (DAPI), DiI, Fluorescein isothiocyanate (FITC), and Cell Counting Kit-8 (CCK-8) assay were obtained from Beyotime Technology. (China). Chongqing East Chemical Industry Ltd., Co. (Chongqing, China) provided the chloroform (CHCl3). It was obtained from Avanti Polar Lipids in Alabama (USA) that DC-cholesterol hydrochloride (DC-CHOL), DSPE-PEG 2000-Amine, and DPPC were manufactured. IR780, perfluorohexane (PFH), and Carboplatin (CBP) were purchased from Aladdin Co., Ltd.
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6

Lipid Labeling for Fluorescence Imaging

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The phospholipids 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[amino(polyethylene glycol)-2000] (ammonium salt) (DSPE-PEG-2000-Amine), and 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[biotinyl(polyethylene glycol)-2000] (ammonium salt) (DSPE-PEG-2000-Biotin) were purchased from Avanti Polar Lipids (Merck, Kenilworth, NJ, USA), while Fluorescein-avidin conjugate (FITC-avidin) was purchased from Thermo Fisher Scientific ( Waltham, MA, USA). Dulbecco’s phosphate-buffered saline (DPBS) and chloroform were purchased from Sigma Aldrich (c St. Louis, MO, USA).
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7

Preparation and Characterization of Cationic Micelles

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The micellar carriers were prepared as described previously [12 ]. Briefly, DSPE-mPEG(2000) or DSPE-PEG(2000)-amine (Avanti Polar Lipids) (Suppl. Figure 1) were mixed with 1,1'-dioctadecyl-3,3,3',3' tetramethylindodi-carbocyanine, 4-Chlorobenzene-sulfonate Salt) at a molar ratio of 98/2. The pegylated lipid mixture was then dried down in a glass tube to a thin PEG-lipid film using a rotary evaporator. The lipid film was rehydrated in phosphate buffered saline (PBS) and sonicated for 10 minutes at room temperature until all components were dispersed. The particle sizes of the cationizable, amine-functionalized micelles DSPE-PEG(2000)-amine and methoxy-PEG-DSPE (mPEG-DSPE were measured by dynamic light scattering (DLS) using the Nanobrook Omni (Brookhaven). To determine the effect of pH on zeta potential, the micelles were dispersed in PBS at pH 7.4 and in 20 mM of 2-(N-morpholino) ethanesulfonic acid (MES) buffer at pH 6.5, 6.0, and 5.5 [13 (link)]. Measurements were taken three times per sample and are represented as mean ± standard deviation.
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8

Synthesis of Lipidated Pepstatin A

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1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[amino(polyethylene glycol)-2000] (DSPE-PEG(2000) amine) (Avanti Polar Lipids, Alabaster, AL, USA, 14.0 mg, 5 µmol) was added to a mixture of pepstatin A (Sigma-Aldrich, St. Louis, MO, USA, 6.9 mg, 10 µmol), EDCI (Acros Organics, Geel, Belgium, 3.8 mg, 20 µmol), HOBt (Sigma-Aldrich, 2.0 mg, 15 µmol), and CHCl3 (1.5 mL), which was stirred at 0 °C. The reaction mixture was then left to warm up to approximately 21–23 °C, and this was followed by stirring for 24 h (monitored via UPLC, Waters AQUITY H-Class, water-acetonitrile gradient, RP C18 column). The reaction mixture was then diluted with CHCl3 (30 mL) and washed three times with 5% NaHCO3(aq) and brine. The organic layer was dried over anhydrous Na2SO4 and filtered, and the solvent was evaporated under reduced pressure. The solid obtained was dissolved in acetonitrile/water (1:1 v/v) and lyophilized to obtain lipidated pepstatin A as a colorless powder.
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9

Synthesis and Characterization of MHI-148 Dye Conjugate

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MHI-148: (2-[2-[2-chloro-3-[2-[1,3-dihydro-3,3-dimethyl-1-(5-carboxypentyl)-2H-indol-2-ylidene]-ethylidene]-1-cyclohexene-1-yl]-ethyl]-3,3-dimethyl-1-(5-carboxypentyl)-3H-indolium bromide was purchased from Bioacts (DKC corporation, Incheon, South Korea). DSPE-PEG2000-Amine from Avanti polar lipids (Alabaster, USA). 1-Ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC), N-hydroxysuccinimide (NHS) and dicyclohexylcarbodiimide (DCC) were purchased from Sigma Aldrich, St Louis, USA. Dulbecco’s modified Eagle’s medium (DMEM) and RPMI-1640 were purchased from Thermo Scientific, Waltham, USA. 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) (MTS) was purchased from Promega, Madison, USA. All other reagents were of analytical or chromatographic grade.
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10

Synthesis and Characterization of Dox/ONBs

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All lipids, namely, 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[amino(polyethylene glycol)-2000] (DSPE-PEG-2000-Amine), and 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[biotinyl(polyethylene glycol)-2000] (DSPE-PEG-2000-Biotin) were purchased from Avanti Polar Lipids (Alabaster, AL, USA). An OxiSelect intracellular ROS assay kit for the fluorometric detection of ROS was purchased from Cell Biolabs Inc. (San Diego, CA, USA). A 99.995% pure oxygen cylinder (Daesung gas, Siheung-si, Gyeonggi-do, South, Korea) was used for oxygen supply during the synthesis of Dox/ONBs. Doxorubicin-HCL, chloroform, cell culture media, and other reagents were purchased from Sigma-Aldrich (St. Louis, MO, USA). DoxovesTM (Liposomal Doxorubicin-HCL) was purchased from FormuMax Scientific (Sunnyvale, CA, USA).
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