Generic liposome reagent for immunoassays.
We have derivatized liposomes with antibodies by using avidin to crosslink biotinylated phospholipid molecules in the liposome membranes with biotinylated antibody molecules. A comparison of the biotin binding activity of avidin in solution and avidin associated with liposomes shows that avidin bound to biotinylated phospholipid in liposome membranes retains full binding activity for additional biotin molecules. Changes in the fluorescence spectrum of avidin have been used to characterize the binding capacity of avidin for biotin in solution, and change in intensity of light scattered due to aggregation of liposomes was used to measure the biotin binding activity of avidin associated with liposomes. Relative amounts of the biotinylated phospholipid, avidin, and biotinylated antibody have been optimized to produce stable liposomes which are derivatized with up to 1.7 nmol of antibody/mumol of lipid. These derivatized liposomes are highly reactive to immunospecific aggregation in the presence of multivalent antigen. A linear increase in light scattering was recorded between 1 and 10 pmol of antigen. This work shows that liposomes containing biotinylated phospholipid can be a successful generic reagent for immunoassays.
Plant
A L
AL
National Institute of Standards and Technology, Center for Analytical Chemistry, Gaithersburg, Maryland 20899.
Brizgys
M V
MV
Locasio-Brown
L
L
Durst
R A
RA
D000906:Antibodies / Q000276:immunology; D000915:Antibody Affinity; D001360:Avidin; D001710:Biotin; D002851:Chromatography, High Pressure Liquid; D003432:Cross-Linking Reagents; D004337:Drug Carriers; D004797:Enzyme-Linked Immunosorbent Assay; D008081:Liposomes; D013050:Spectrometry, Fluorescence
D000906:Antibodies; D003432:Cross-Linking Reagents; D004337:Drug Carriers; D008081:Liposomes; D001360:Avidin; D001710:Biotin