@article {23,
	title = {PEG-PE/phosphatidylcholine mixed immunomicelles specifically deliver encapsulated taxol to tumor cells of different origin and promote their efficient killing.},
	journal = {J Drug Target},
	volume = {11},
	year = {2003},
	month = {2003 Feb},
	pages = {87-92},
	abstract = {<p>Mixed micelles were prepared from poly(ethyleneglycol)-distearyl phosphoethanolamine (PEG2000-PE) and egg phosphatidylcholine. The micelles were covalently modified with the nucleosome-specific monoclonal antibody 2C5 known to recognize and bind a variety of tumor cells via their surface-bound nucleosomes. Covalent attachment of 2C5 antibody was performed via a micelle-incorporated PEG-PE with the distal terminus of the PEG block activated with p-nitrophenylcarbonyl group (pNP-PEG-PE). Micelle surface-attached 2C5 antibody maintained its specific activity. 2C5-targeted immunomicelles were able to carry more than 3 wt\% of taxol. Taxol-loaded immunomicelles specifically recognized tumor cell lines of several types. The cytotoxicity of 2C5-targeted taxol-loaded immunomicelles in a cell culture model was much higher when compared with free taxol or taxol in non-targeted micelles.</p>},
	keywords = {Animals, Antibodies, Monoclonal, Antineoplastic Agents, Cell Survival, Chromatography, High Pressure Liquid, Drug Carriers, Electrophoresis, Polyacrylamide Gel, Mice, Micelles, Nucleosomes, Paclitaxel, Particle Size, Phosphatidylcholines, Phosphatidylethanolamines, Polyethylene Glycols, Tumor Cells, Cultured},
	issn = {1061-186X},
	doi = {10.1080/1061186031000138623},
	author = {Gao, Z and Lukyanov, A N and Chakilam, A R and Torchilin, V P}
}
@article {30,
	title = {p-Nitrophenylcarbonyl-PEG-PE-liposomes: fast and simple attachment of specific ligands, including monoclonal antibodies, to distal ends of PEG chains via p-nitrophenylcarbonyl groups.},
	journal = {Biochim Biophys Acta},
	volume = {1511},
	year = {2001},
	month = {2001 Apr 2},
	pages = {397-411},
	abstract = {<p>We have attempted to simplify the procedure for coupling various ligands to distal ends of liposome-grafted polyethylene glycol (PEG) chains and to make it applicable for single-step binding of a large variety of a primary amino group-containing substances, including proteins and small molecules. With this in mind, we have introduced a new amphiphilic PEG derivative, p-nitrophenylcarbonyl-PEG-1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (pNP-PEG-DOPE), synthesized by reaction of DOPE with excess of bis(p-nitrophenylcarbonyl)-PEG in a chloroform/triethylamine mixture. pNP-PEG-DOPE readily incorporates into liposomes via its PE residue, and easily binds primary amino group-containing ligands via its water-exposed pNP groups, forming stable and non-toxic urethane (carbamate) bonds. The reaction between the pNP group and the ligand amino group proceeds easily and quantitatively at pH around 8.0, and remaining free pNP groups are promptly eliminated by spontaneous hydrolysis. Therefore, pNP-PEG-DOPE could serve as a very convenient tool for protein attachment to the distal ends of liposome-grafted PEG chains. To investigate the applicability of the suggested protocol for the preparation of long-circulating targeted liposomes, we have coupled several proteins, such as concanavalin A (ConA), wheat germ agglutinin (WGA), avidin, monoclonal antimyosin antibody 2G4 (mon2G4), and monoclonal antinucleosome antibody 2C5 (mon2C5) to PEG-liposomes via terminal pNP groups and studied whether the specific activity of these immobilized proteins is preserved. The method permits the binding of several dozens protein molecules per single 200 nm liposome. All bound proteins completely preserve their specific activity. Lectin-liposomes are agglutinated by the appropriate polyvalent substrates (mannan for ConA-liposomes and glycophorin for WGA-liposomes); avidin-liposomes specifically bind with biotin-agarose; antibody-liposomes demonstrate high specific binding to the substrate monolayer both in the direct binding assay and in ELISA. A comparison of the suggested method with the method of direct membrane incorporation was made. The effect of the concentration of liposome-grafted PEG on the preservation of specific protein activity in different coupling protocols was also investigated. It was also shown that pNP-PEG-DOPE-liposomes with and without attached ligands demonstrate increased stability in mouse serum.</p>},
	keywords = {Animals, Antibodies, Monoclonal, Avidin, Drug Stability, Hydrogen-Ion Concentration, Lectins, Ligands, Liposomes, Mice, Models, Chemical, Nitro Compounds, Phosphatidylethanolamines, Polyethylene Glycols, Protein Binding, Proteins, Surface-Active Agents},
	issn = {0006-3002},
	author = {Torchilin, V P and Levchenko, T S and Lukyanov, A N and Khaw, B A and Klibanov, A L and Rammohan, R and Samokhin, G P and Whiteman, K R}
}
