Egg phosphatidylcholine

Manufactured by Lipoid

Sourced in Germany, United States

Egg phosphatidylcholine is a naturally occurring phospholipid obtained from egg yolks. It is a key component of cell membranes and plays a role in maintaining cellular structure and function.

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Phosphatidylcholine (14 citations) Egg phosphatidylcholine (EPC) (8 citations) Egg yolk phosphatidylcholine (5 citations) Egg phosphatidylcholine (egg PC) (3 citations) Egg phosphatidylcholine (PC) (2 citations) Egg phosphatidylcholine (Phospholipon 90G) (2 citations)

13 protocols using egg phosphatidylcholine

Targeted Lipid Nanoparticle Preparation

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Egg phosphatidylcholine was supplied by Lipoid GmbH (Ludwigshafen, Germany). DSPE-PEG₂₀₀₀ (DSPE-PEG) and DSPE-PEG₂₀₀₀-NHS (DSPE-PEG-NHS) were obtained from NOF Corporation (Tokyo, Japan). Coumarin-6, cholesterol, sulforhodamine B, and tris base were bought from Sigma-Aldrich (St Louis, MO, USA). The Ac-RRK(HHHHHHH) RR-NH₂ peptide (H₇K(R₂)₂) was synthesized by Beijing Scilight Biotechnology Co., Ltd. (Beijing, China). DSPE-PEG-H₇K(R₂)₂ was synthesized by our laboratory according to a previously reported method.23 (link),24 (link) PTX was obtained from Mei-Lian Co., Ltd. (Chongqing, China). Near-infrared lipophilic carbocyanine dye 1,10-dioctadecyltetramethyl indotricarbocyanine iodide (DiR) and Coumarin-6 were supplied by Biotium, Inc. (Hayward, CA, USA).
SPIO NPs were synthesized by our lab according to the reported reference.25
The human breast carcinoma MDA-MB-231 cell line was obtained from the Chinese Academy of Sciences Cell Bank (Shanghai, China). This cell line was cultured according to the recommended instructions.

Zheng X.C., Ren W., Zhang S., Zhong T., Duan X.C., Yin Y.F., Xu M.Q., Hao Y.L., Li Z.T., Li H., Liu M., Li Z.Y, & Zhang X. (2018). The theranostic efficiency of tumor-specific, pH-responsive, peptide-modified, liposome-containing paclitaxel and superparamagnetic iron oxide nanoparticles. International Journal of Nanomedicine, 13, 1495-1504.

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Synthesis and Characterization of Doxorubicin-Loaded Nanoparticles

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Sodium citrate (>99%), cobalt chloride hexahydrate (99.99%), sodium borohydride (99%), and chloroauric acid trihydrate (American Chemical Society reagent grade) were purchased from Thermo Fisher Scientific (Waltham, MA) and were used as received. Octadecyl-3-Mercaptopionate (OMP) was from Chemical Indoustry Co.(Japan). Doxorubicin.HCl (DOX) was gifted from Zhejiang Hisun Pharmaceutical Co, Ltd, (Taizhou Zhejiang, China). 1,2-dipalmitoyl-sn-glycero-3-phosphatidylcholine (DPPC), Egg phosphatidylcholine (EPC), Dipalmitoylphosphatidylcholine (DPPC), Hydrogenated soybean phosphatidylcholine (HSPC), cholesterol (Chol), and 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethyleneglycol)-2000] (DSPE-PEG₂₀₀₀) were purchased from Lipoid (Germany). 1,1’-dioctadecyl-3,3,3,3’-tetramethylindodicarbocyanine, 4-chlorobenzenesulfonate salt (DiD), 1,1'-Dioctadecyl-3,3,3',3'-Tetramethylindotricarbocyanine Iodide (DiR) were obtained from Invitrogen (Carlsbad, CA, USA). 3-(4,5-dimethylthiazol-2-yl)-2,5 -diphenyltetrazolium bromide (MTT) were purchased from Sigma (St Louis, MO). All other solvents were of analytical or chromatographic grade.

You J., Zhang P., Hu F., Du Y., Yuan H., Zhu J., Wang Z., Zhou J, & Li C. (2014). Near-infrared light sensitive liposomes for the enhanced photothermal tumor treatment by the combination with chemotherapy. Pharmaceutical research, 31(3), 554-565.

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Clodronate-Containing Liposomes for Macrophage Depletion

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Clodronate-containing liposomes were prepared as described by van Rijn et al. [26 (link)]. Briefly, a mixture of egg-phosphatidylcholine, egg-phosphatidylglycerol (both from Lipoid, Ludwigshafen, Germany), and cholesterol (Sigma, St. Louis, MO) was dissolved in ethanol in a molar ratio of 10 : 1 : 1.5 and evaporated to dryness by rotation under reduced pressure. The lipid film was hydrated in an aqueous solution containing clodronate (Cl2MDP, concentration 60 mg/mL, Bonefos; Schering, Weesp, Netherlands). The unencapsulated clodronate was removed by repeated washing using PBS pH 7.4 and ultracentrifugation (Beckman Optima LE-80 K; Palo Alto, CA) at 200,000 g for 30 min. After washing, the pellet was resuspended in PBS at a concentration of 90 mM phospholipids. The final concentration of clodronate was determined using spectrophotometry at a wavelength of 238 nm after extraction and binding to Cu⁺ and appeared to be within a range of 2-2.5 mg/mL. Mice prone for depletion of macrophages received 0.2 mL of the liposome-suspension intravenously one day before injection of huPBMCs.

Hogenes M.C., van Dorp S., van Kuik J., Monteiro F.R., ter Hoeve N., Guedes L., van Dijk M.R., Martens A.C, & de Weger R.A. (2019). Modifying Graft-versus-Host Disease in a Humanized Mouse Model by Targeting Macrophages or B-Cells. Journal of Immunology Research, 2019, 3538963.

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Nano-Encapsulated Drug Delivery System

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Sodium alginate (CP, viscosity: 200 ± 20 mPa·s) was purchased from Shanghai Macklin Biochemical Co., Ltd. (Shanghai, China). Egg Phosphatidyl Choline (EPC) was purchased from Lipoid GmbH (Ludwigshafen, Germany). Fluorescein sodium, CCK-8 kit, and DIR were purchased from Yeasen Biotechnology (Shanghai) Co., Ltd. (Shanghai, China). Mannitol was acquired from Shanghai Aladdin Biochemical Technology Co., Ltd. (Shanghai, China). Anhydrous calcium chloride, sodium dodecyl sulfate, and dichloromethane were purchased from Anhui Senrise Technology Co., Ltd. (Shanghai, China). Poly lactic-co-glycolic acid (molecular weight 24,000–38,000) was purchased from Xi’an ruixi Biological Technology Co., Ltd. (Xi’an, China). Methanol, absolute ethanol, and acetonitrile were purchased from Thermo Fisher Scientific (Shanghai, China). Fetal bovine serum (FBS) was purchased from Gibco (Grand Island, NY, USA). Antibiotics (100 U/mL penicillin and 100 U/mL streptomycin) and non-essential amino acids (NEAA) were purchased from Sigma Aldrich (St. Louis, MO, USA). All other chemicals used in this study were of analytical or high-performance liquid chromatography (HPLC) grade.

Zhang X., Zhu R., Wang X., Wang H., Xu Z., Wang Y., Quan D, & Shen L. (2023). Core-Shell Microspheres Prepared Using Coaxial Electrostatic Spray for Local Chemotherapy of Solid Tumors. Pharmaceutics, 16(1), 45.

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Formulation and Characterization of Azithromycin-Roxithromycin Nanoparticles

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Azithromycin was gifted by Sichuan Jiangchuan Pharmaceutical Co., Ltd. (Sichuan, China). Roxithromycin was obtained from Dalian Meilun Biological Technology Co., Ltd. (Liaoning, China). Medium-chain triglycerides were provided by Tieling North Asia Pharmaceutical Co., Ltd. (Liaoning, China). Egg phosphatidylcholine came from Lipoid E80 (Ludwigshafen, Germany). Distilled water was self-prepared. The solvents used for the mobile phases were high performance liquid chromatography grade. All other chemicals were of analytical grade.

Huang R., Zhang Y., Wang T., Shen L., Zhang Z., Wang Y, & Quan D. (2018). Creation of an assessment system for measuring the bitterness of azithromycin-containing reverse micelles. Asian Journal of Pharmaceutical Sciences, 13(4), 343-352.

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Nanobody-based Protein Labeling Protocol

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MicroBCA Protein Assay Kit, AlexaFluor 488 NHS ester, CellTracker Green CMFDA dye, CellTracker Deep Red dye, CellTracker Red CMTPX dye and the pcDNA3.1 vector with Neomycin resistance were purchased from Thermo Fisher Scientific (Waltham, USA). Sepharose CL-4B and cholesterol were obtained from Sigma-Aldrich (Steinheim, Germany) and TALON Superflow Metal Affinity Resin was from Clontech Laboratories, Inc. (Saint-Germain-en-Laye, France). Phospholipids were purchased from Avanti Polar Lipids (Birmingham, USA), except for egg phosphatidylcholine, which was obtained from Lipoid AG (Steinhausen, Switzerland). Cell culture flasks were obtained from Greiner Bio-One (Alphen aan de Rijn, The Netherlands). pET28a-EGa1 and pAX51-R2 vectors encoding EGa1 (PDB ID: 4KRN) and R2 (PDB ID: ; 1QD0) Myc-tagged nanobodies, respectively, were kindly provided by Dr S. Oliveira (Department of Biology, Utrecht University, Utrecht, The Netherlands).

Kooijmans S.A., Gitz-Francois J.J., Schiffelers R.M, & Vader P. (2018). Recombinant phosphatidylserine-binding nanobodies for targeting of extracellular vesicles to tumor cells: a plug-and-play approach †Electronic supplementary information (ESI) available. See DOI: 10.1039/c7nr06966a. Nanoscale, 10(5), 2413-2426.

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Synthesis and Characterization of iRGD-Targeted CLA-PTX Nanoparticles

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PTX was obtained from Yunnan Hande Bio-Tech (Kunming, People’s Republic of China [PRC]). CLA-PTX was synthesized from CLA and PTX according to our previous report.21 (link) The structure of CLA-PTX is shown in Figure 1. The iRGD peptide (CRGDK/RGPD/EC) was supplied by GL Bio-Chem (Shanghai, PRC). Cholesterol, sulforhodamine B, chlorpromazine, amiloride, filipin, methyl-β-cyclodextrin (M-β-CD) and Tris base were purchased from Sigma-Aldrich (St Louis, MO, USA). Coumarin-6 and Hoechst 33258 were obtained from Thermo Fisher Scientific (Waltham, MA, USA). Egg phosphatidylcholine was purchased from Lipoid (Ludwigshafen, Germany). 1,2-distearoyl-sn-glycero-3-phosphoethanolamine- N-[methoxy(polyethylene glycol)-2000] (DSPE-PEG) was obtained from Avanti Polar Lipids (Alabaster, AL, USA). iRGD-PEG-DSPE, as shown in Figure 1, was synthesized according to our previously reported method.26 (link)
Cell-culture media, Dulbecco’s Modified Eagle’s Medium DMEM (high glucose), penicillin–streptomycin, fetal bovine serum, and L-glutamine were purchased from Thermo Fisher Scientific. All solvents and other reagents were of analytical grade.

Du R., Zhong T., Zhang W.Q., Song P., Song W.D., Zhao Y., Wang C., Tang Y.Q., Zhang X, & Zhang Q. (2014). Antitumor effect of iRGD-modified liposomes containing conjugated linoleic acid–paclitaxel (CLA-PTX) on B16-F10 melanoma. International Journal of Nanomedicine, 9, 3091-3105.

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Paclitaxel Oleate Nanoparticle Preparation

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The LDE-paclitaxel oleate preparation was made according to the methods described in
previous studies (11 (link),27 (link),28 (link)). Briefly, a lipid
mixture composed by 135 mg cholesteryl oleate (Alfa Aesar, USA), 333 mg egg
phosphatidylcholine (Lipoid, Germany), 132 mg miglyol 812N (Sasol Germany GmbH,
Germany), 6 mg cholesterol (Fabrichem, USA) and 60 mg of paclitaxel oleate
(Pharmaceuticals Co., China), was added to an aqueous phase consisting of 100 mg of
polysorbate 80 (Merck, Germany) and 10 mL Tris-HCl buffer, pH 8.05. A pre-emulsion
was obtained by ultrasonic radiation until complete solubilization of the drug.
Emulsification of all lipids, drug and aqueous phase was obtained by high-pressure
homogenization using an Emulsiflex C5 homogenizer (Avestin, Canada). After
homogenization cycles, the formed emulsion was centrifuged and the nanoparticle
sterilized by passage through 0.22 μm pore polycarbonate filter (Millipore, USA) and
kept at 4°C until used.

Freitas S.C., Tavares E.R., Silva B.M., Meneghini B.C., Kalil-Filho R, & Maranhão R.C. (2018). Lipid core nanoparticles resembling low-density lipoprotein and regression of atherosclerotic lesions: effects of particle size. Brazilian Journal of Medical and Biological Research, 51(3), e7090.

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Lipid-Based Methotrexate Nanoemulsion Preparation

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LDE-MTX was prepared from a lipid mixture composed of 100 mg cholesteryl oleate, 200 mg egg phosphatidylcholine (Lipoid, Germany), 10 mg triglycerides, 12 mg cholesterol and 60 mg of MTX (5 (link),19 (link)). The aqueous phase (100 mg of polysorbate 80 and 10 mL of Tris-HCl buffer, pH 8.05) was kept at room temperature. The pre-emulsion was obtained adding the hydrophilic phase to the oil phase by ultrasonic radiation until complete dissolution of the drug. Emulsification of the compounds was obtained by high-pressure homogenization using an Emulsiflex C5 homogenizer (Avestin, Canada). After homogenization at constant temperature, the nanoemulsion was centrifuged at 1800 g for 15 min at 4°C to separate the emulsified from unbound MTX. The nanoemulsion was sterilized by passage through 0.22 μm pore filter (Millipore, USA) and kept at 4°C until it was used. The association of MTX to LDE was measured by HPLC before the treatment (22 (link)).

Fiorelli A.I., Lourenço-Filho D.D., Tavares E.R., Carvalho P.O., Marques A.F., Gutierrez P.S., Maranhão R.C, & Stolf N.A. (2017). Methotrexate associated to lipid core nanoparticles improves cardiac allograft vasculopathy and the inflammatory profile in a rabbit heart graft model. Brazilian Journal of Medical and Biological Research, 50(10), e6225.

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Optimized Lipid-based Nanocarrier Formulation

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L-serine was purchased from Sigma-Aldrich Company (St. Louis, MO, USA). Egg phosphatidylcholine was obtained from Lipoid GmbH (Ludwigshafen, Germany). Carbopol ETD 2020 (Acrylates/C10-30 Alkyl Acrylate Crosspolymer, International Nomenclature of Cosmetic Ingredients, INCI name) was provided by The Lubrizol Corporation (Wickliffe, OH, USA) and Rhesperse RM100 (Sodium Polyacrylate, INCI name) was supplied from Sensient Cosmetic Technologies (South Plainfield, NJ, USA). All other chemicals were reagents grade and obtained from Sun Fine Global Co., Ltd. (Cheongju-si, Chungbuk, Korea), Ewoo Chemicals (Seoul, Korea), or the KCC Corporation (Seoul, Korea).

Kim H., Ro J., Barua S., Hwang D.S., Na S.J., Lee H.S., Jeong J.H., Woo S., Kim H., Hong B., Yun G., Kim J.H., Yoon Y.H., Park M.G., Kim J., Sohn U.D, & Lee J. (2015). Combined Skin Moisturization of Liposomal Serine Incorporated in Hydrogels Prepared with Carbopol ETD 2020, Rhesperse RM 100 and Hyaluronic Acid. The Korean Journal of Physiology & Pharmacology : Official Journal of the Korean Physiological Society and the Korean Society of Pharmacology, 19(6), 543-547.

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