• Journal of Pharmaceutical Investigation
• /
• v.21 no.3
• /
• pp.161-170
• /
• 1991

The storage stabilities of immunoliposomes incorporated with variable amounts of ganglioside $G_{M1}$ were investigated as a function of time. temperature. and composition by observing absorbance of visible light and calcein release. In the column chromatographe, the layer of unsaturated PE(dioleoylphosphatidylethanolamine : DOPE). unable to form stable liposomes at physiological temperature and pH, were formed when palmitoyl-immunoglobulin G(IgG) $(2.5{\times}10^{-4}\;mol/DOPE\;mol)$ added. The incorporation of ganglioside $G_{M1}$ into immunoliposome. enhanced the stabilities of bilayers during the extended period of storage. The turbidities of immunoliposomes coated with ganglioside $G_{M1}$ exhibited the maximum near 20 mol% $G_{M1}/DOPE$ mol. probably because of the disturbance of the bilayer characteristics, i.e., layer transition or reorientation of interaction sites. At low temperature. the higher stability was achieved than at elevated temperatures. After one week of storage. the redispersed liposomal solutions at lower temperatures maintained the original elution patterns in chromatography but broader distribution at elevated temperatures. During the storage, it is suggested the aggregation is the more dominant phenomena for liposomes kept at $5^{\circ}C$ than the fusion. while he fusion is at elevated temperatures.

• Journal of Microbiology and Biotechnology
• /
• v.18 no.10
• /
• pp.1689-1694
• /
• 2008

The ability to detect Salmonella spp. is essential in the prevention of foodborne illness. This study examined a Salmonella spp. detection method involving the application of immunomagnetic separation and immunoliposomes (IMS/IL) encapsulating sulforhodamine B (SRB), a fluorescent dye. A quantitative assay was conducted by measuring the fluorescence intensity of SRB that was produced from an immunomagnetic bead-Salmonella spp.-immunoliposome complex. The results indicated detection limits of $2.7{\times}10^{5}$ and $5.2{\times}10^{3}$ CFU/ml for Salmonella enterica subsp. enterica serovar Enteritidis (S. Enteritidis) and Salmonella enterka subsp. enterka serovar Typhimurium (S. Typhimurium), respectivley. The signal/noise ratio was improved by using 4% skim milk as a wash solution rather than 2% BSA. In addition, higher fluorescence intensity was obtained by increasing the liposome size. Compared with the conventional plating method, which takes 3-4 days for the isolation and identification of Salmonella spp., the total assay time of to h only including 6 h of culture enrichment was necessary for the Salmonella detection by IMS/IL. These results indicate that the IMS/ IL has great potential as an alternative rapid method for Salmonella detection.

• 한국생물공학회:학술대회논문집
• /
• 2000.11a
• /
• pp.206-207
• /
• 2000

A disposable, electro-chemiluminescent immunosensor utilizing a screen-printed carbon electrode and liposome coupled to antibody as tracer has been constructed. In proportion to the analyte (Legionella species as a model) concentration, the analyte-immunoliposome complexes were transferred by the capillary action through a membrane strip to the electrode, the liposomes were lysed in the presence of detergent, and ruthenium was released for signal generation. Such performance of the immunosensor was appropriate for a point-of-care testing.

• BMB Reports
• /
• v.31 no.1
• /
• pp.95-100
• /
• 1998

Two innovative methods to prepare target-sensitive immunoliposomes containing doxorubicin by coupling monoclonal antibodies (mAb DH2, SH1) specific to cancer cell surface antigens ($G_{M3}$, $Le^X$) have been developed and are described here. Firstly, liposomes containing N-glutaryl phosphatidylethanolamine (NGPE) were prepared, followed by the encapsulation of doxorubicin, DH2 or SH1 antibodies were conjugated to NGPE in the liposomes (direct coupling). Secondly, liposomes were prepared with NGPE/mAb conjugates by the detergent dialysis method (conjugate insertion), and then doxorubicin was encapsulated by proton gradient. The immunoliposomes prepared by both methods were able to specifically bind to the surface of the tumor cells - B16BL6 mouse melanoma cells. The efficiencies of doxorubicin-entrapping into liposomes prepared by direct coupling and conjugate insertion was about 98% and 25%, respectively. These types of liposomal formulation are sensitive to target cells, which can be useful for various clinical applications.

• Journal of Pharmaceutical Investigation
• /
• v.22 no.2
• /
• pp.115-124
• /
• 1992

The characteristics and stabilities of phosphatidylcholine liposomes covalently coupled with immunoglobulin fragments prepared by the REV method were investigated by the dynamic light scattering, absorbance and calcein release. Using a sulfhydryl-reactive phospholipid derivative of N-[4$({\rho}-maleimido-phenyl)$ butyl] phosphatidylethanolamine (MPB-PE), Fab' antibody fragments were covalently combined with preformed large unilamellar vesicles (LUV), Coupling ratio was $250\;{\mu}g$ of $Fab'/{\mu}mol$ of phospholipid in vesicles, From dynamic light scattering, it was found that the size of the vesicles increases as the ratio of cholesterol to lipid increases, but that apparently, the size of liposomes was not sensitive to the existence of Fab' fragments. Regardless of inserting Fab' fragments, the absorbance of liposomes decreased as the amounts of bile salt (BS) added. At very low BS concentrations, BS/lipid aggregates would be formed in the outer vesicles monolayer, while, at the high BS concentrations, mixed micelles would be preferred. The vesicles incorporated with Fab' fragments, however, are more resistant to the bile salts than the MPB-PE vesicle are. The absorbance of vacant liposomes and calcein release resulted in that the Fab' vesicles and MPB-PE vesicles by the REV method are very stable, but that those by the sonication method sufferred the significant change of turbidities.

• Archives of Pharmacal Research
• /
• v.27 no.12
• /
• pp.1263-1269
• /
• 2004

The objective of this study was to characterize immunoliposomes carrying plasmid DNA with optimal encapsulation efficiency and antibody density. Plasmid DNA was encapsulated by the freezing/thawing method into liposomes composed of POPC (1-palmitoyl-2-oleoyl-sn-glycerol- 3-phosphocholine), DDAB (didodecyl dimethyl ammonium bromide), DSPE-PEG 2000 (distearoyl phosphatidyl ethanolamine polyethylene glycol 2000) and DSPE-PEG 2000-maleimide. The liposomes carrying plasmid DNA were extruded through two stacked polycarbonate filters, of different pore size, to control the liposome size. Then, rat IgG molecules were conjugated to the liposomes. The immunoliposomes containing plasmid DNA were separated from the free plasmid DNA and unconjugated IgG by Sepharose CL-4B column chromatography. The DNA amount encapsulated was affected by DDAB (cationic lipid) concentration, the initial amount of plasmid DNA between 10 ${\mu}g$ and 200 ${\mu}g$, the total lipid amount and plasmid DNA size, but not significantly by liposome size. By varying the ratio of DSPE-PEG 2000-maleimide to IgG, the number of IgG molecules per liposome was changed significantly.

• Journal of Pharmaceutical Investigation
• /
• v.20 no.3
• /
• pp.135-144
• /
• 1990

The effects of bile salts (BS) on the stability of dioleoylphosphatidylethanolamine (DOPE) liposomes were investigated, observing apparent absorbance of vacant liposomes and calcein release from entrapped liposomes. Unilamellar liposomes were prepared by using a small quantity of palmitoly-immunoglobulin G(IgG) ($2.5{\times}10^{-4}$ mo1/lipid mol) to stabilize the bilayer phase of the unsaturated DOPE which by itself does not form stable liposomes. The destabilization of PE immunoliposomes by papain, clearly demonstrates that the IgG is essential for stabilization of PE bilayer. Approximately 4% of the entrapped calcein was released from the PE liposomes after 1 hr from liposome formation. Calcein release and absorbance of liposomes depended on the BS/lipid ratio because of the solubilization of lipid molecule in bilayer and the formation of mixed micelles. At very low BS concentrations, the incorporation of BS induced BS/lipid aggregates in the outer vesicles monolayer, while high BS concentrations, mixed micelles were formed. Chelate and its conjugates as $3{\alpha},\;7{\alpha},\;12{\alpha}-trihydroxy$ BS induce the concentration of the $3{\alpha}$, $12{\alpha}-dihydroxy$ BS at half-maximal solubilization of immunoliposomes to approximately 2.5-, or 5-fold. Conjugation of BS with glycine or taurine slightly enhanced their capacities to perturb membranes.

• Journal of Microbiology and Biotechnology
• /
• v.26 no.11
• /
• pp.1855-1862
• /
• 2016

Cronobacter sakazakii (C. sakazakii) is a foodborne pathogen, posing a high risk of disease to infants and immunocompromised individuals. In order to develop a quick, easy, and sensitive assay for detecting C. sakazakii, a rabbit anti-C. sakazakii immunoglobulin G (IgG) was developed using sonicated cell protein from C. sakazakii. The developed anti-C. sakazakii (IgG) was of good quality and purity, as well as species-specific. The developed rabbit anti-C. sakazakii IgG was attached to the surface of a sulforhodamine B-encapsulated liposome to form an immunoliposome. A test strip was then prepared by coating goat anti-rabbit IgG onto the control line and rabbit anti-C. sakazakii IgG onto the test line, respectively, of a plastic-backed nitrocellulose membrane. A purple color signal both on the test line and the control line indicated the presence of C. sakazakii in the sample, whereas purple color only on the control line indicated the absence of C. sakazakii in the sample. This immunochromatographic strip assay could produce results in 15 min with a limit of detection of $10^7CFU/ml$ in C. sakazakii culture. The immunochromatographic strip assay also showed very good specificity without cross-reactivity with other tested Cronobacter species. Based on these results, the developed immunochromatographic strip assay is efficient for the detection of C. sakazakii and has high potential for on-site detection.