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4 Cellular and Molecular Biology Letters

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Preparation of PEG-grafted immunomagneto-liposomes and their application in cell sorting

100%

Domingo J. C. , Mercadal M. , De Madariaga M. A.

Cellular and Molecular Biology Letters

1999

tom 04

nr 2

Surface modified liposomes by coating with charged hydrophilic molecules

100%

Sagrista M. L. , Mora M. , De Madariaga M. A.

Cellular and Molecular Biology Letters

1999

tom 04

nr 2

Surface modified liposomes by coating with charged hydrophilic molecules

100%

Sagrista M. L. , Mora M. , De Madariaga M. A.

Cellular and Molecular Biology Letters

2000

tom 05

nr 1

The design of liposomes with a hydrophilic/steric barrier at their bilayer surface allows the modification of their pharmacokinetics and reduces the uptake by the RES. Liposomes can be coated by hydrophilic molecules such as polysaccharides, which disguise the vesicle surface by creating a three-dimensional matrix near them and prevent the binding of plasma proteins and their recognition by some cellular receptors. All these considerations, and previous results obtained in our laboratory showing the formation of stable GAG-liposome complexes, have lead us to think about the use of the negatively charged glycosaminoglycans (GAGs), alternately to other molecules such as the monosialoganglioside GM1, more expensive, or polyethylene glycol (PEG-PE) that can disturb the structural organization of the bilayer. The present paper describes the effect of the incorporation of GAGs to phospholipid vesicles, in relation to their electrical and permeability properties. The results obtained show that there is an effective coating of the bilayer surface when glycosaminoglycans are added to liposome suspensions. The shielding of the negative surface charge by the neutral hyaluronic acid, in the absence of calcium, and the increase in the negative charge when the negative polyelectrolytes chondroitin sulfate, heparin or dextran sulfate are added to calcium-containing liposome suspensions account for the formation of stable liposome-GAG complexes. Moreover, the reduced permeability of the GAG-coated liposomes points out on their ability to hold encapsulated drugs and, so, their potential usefulness as drug-sustained release carriers. The hydrophilic coating will give to these liposomal carriers long-circulating properties.

Preparation of PEG-grafted immunomagnetoliposomes and their application in cell sorting

88%

Domingo J. C. , Mercadal M. , Petriz J. , Garcia J. , De Madariaga M. A.

Cellular and Molecular Biology Letters

1999

tom 04

nr 4

Immunomagnetic systems have been used for positive selection of the cell fraction from a mixture using appropiate surface markers with satisfactory results, as hematopoietic CD34+ cells. In this work, we report the development of poly(ethylene glycol) (PEG)-grafted immunoliposomes loaded with dextran-magne-tite particles as the separation vehicles for immunomagnetic separation techniques. The magnetic ferrofluid was encapsulated into PEG-liposomes by the FTS methodology. The magnetoliposomes had a liposomal size around 800 nm and a Fe/lipid molar ratio of 0.87±0.30, and were retained in the magnetic field created by the MiniMACS system. Anti-CD34 immunomagnetoliposomes were prepared by coupling the My10 mAb and bound specifically to CD34+ KG-1a cells in culture and in mixtures with CD34- CHO cells. The magnetic cell sorting was carried out in cell mixtures KG-1a/CHO with a 10% initial of CD34+ KG-1a cells. The purity of the MiniMACS-positive fraction and the capture efficiency depended on the liposome concentration and antibody density used, related to the nonspecific cell binding of immunomagnetoliposomes due to the ferrofluid adsorbed and to the presence of whole antibody molecules in the liposome surface. The CD34+ cells isolated retained viability with an estimated recovery of 45-50%.