Regular paper
Targetability of novel immunoliposomes modified with amphipathic poly(ethylene glycol) s conjugated at their distal terminals to monoclonal antibodies

https://doi.org/10.1016/0005-2736(94)00263-OGet rights and content
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Abstract

Distearoyl-N-(3-carboxypropionoyl poly(ethylene glycol) succinyl)phosphatidylethanolamine (DSPE-PEG-COOH) was newly synthesized and used to prepare novel immunoliposomes carrying monoclonal antibodies at the distal ends of the PEG chains (Type C). Liposomes were prepared from egg phosphatidylcholine (ePC) and cholesterol (CH) (2:1, m/m) containing 6 mol% of DSPE-PEG-COOH, and a monoclonal IgG antibody, 34A, which is highly specific to pulmonary endothelial cells, was conjugated to the carboxyl groups of DSPE-PEG-COOH to give various amounts of antibody molecules per liposome. Other immunoliposomes with PEG coating (Type B) or without PEG coating (an earlier type of immunoliposome, Type A) were prepared for comparison. The average molecular weight of PEG in Type B or C immunoliposomes was 2000. Type B and Type C liposomes without antibodies showed prolonged circulation time and reduced reticulo-endothelial system (RES) uptake owing to the presence of PEG. These three different types of 34A-immunoliposomes with 30–35 antibody molecules per vesicle were injected into mice to test the immunotargetability to the lung. The efficiency of lung binding of 34A-Type B was one-half of that of 34A-Type A, though a large amount of 34A-Type B remained in the blood circulation for a long time, suggesting that the steric hindrance of PEG chains reduced not only the immunospecific antibody-antigen binding, but also the RES uptake. The degree of lung binding of 34A-Type C was about 1.3-fold higher than that of 34A-Type A, indicating that recognition by the antibodies attached to the PEG terminal was not sterically hindered and that the free PEG (i.e., that not carrying antibody) was effective in increasing the blood concentration of immunoliposomes by enabling them to evade RES uptake. The latter phenomenon was confirmed by using nonspecific antibody-Type C immunoliposomes (14-Type C), which showed a high blood level for a long time. Our approach provides a simple means of conjugating antibodies directly to the distal end of PEG which is already bound to the liposome membrane, and should contribute to the development of superior targetable drug delivery vehicles for use in diagnostics and therapy.

Keywords

Liposome
Immunoliposome
Antibody
Poly(ethylene glycol)
Drug delivery system

Abbreviations

CH, cholesterol
DF, deferoxamine
DSPE, distearoyl phosphatidylethanolamine
DSPE-PEG, distearoyl-N-(monomethoxy poly(ethylene glycol) succinyl)phosphatidylethanolamine
DSPE-PEG-COOH, distearoyl-N-(3-carboxypropionoyl poly(ethylene glycol) succinyl)phosphatidylethanolamine
ePC, egg phosphatidylcholine
Mes, 2-(N-morpholino)ethanesulfonic acid hemisodium salt
NGPE, N-glutaryldistearoylphosphatidylethanolamine
PEG-OSu, monomethoxy poly(ethylene glycol) succinimidyl succinate
PEG-2)Su, poly(ethylene glycol) bis(succinimidyl succinate)
SUV, small unilamellar vesicle
34A, rat monoclonal IgG2a antibody
14, rat monoclonal IgG2a antibody, a subtype-matched nonspecific control antibody to 34A

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