Enhanced fluorescence in indirect immunophenotyping by the use of fluorescent liposomes.
Small unilamellar liposomes were optimised for cell phenotyping by indirect immunofluorescence. This involved selection and covalent attachment to the liposome of a polyspecific ligand for cell-bound antibody. For this purpose sheep anti-mouse antibody was preferred to protein A because of its ability to attach to cell-bound IgG1 as well as IgG2 at physiological pH. The maximally fluorescent concentration of encapsulated carboxyfluorescein was determined to be 20 mM and liposomes thus comprised gave up to a nine-fold increase in mean cell fluorescence when compared with sheep anti-mouse antibody conjugated to fluorescein isothiocyanate. There was no parallel increase in background fluorescence. Liposomes retained their targeting and fluorescence properties after 3 months storage. They could be sterilised and were as versatile in use as FITC-antibody conjugates.
Gray
A G
AG
Department of Clinical Haematology, UCMSM, London, U.K.
Morgan
J
J
Linch
D C
DC
Huehns
E R
ER
D000818:Animals; D005434:Flow Cytometry; D016650:Fluorescein-5-isothiocyanate; D005452:Fluoresceins / Q000008:administration & dosage; D005455:Fluorescent Antibody Technique; D006801:Humans; D008081:Liposomes; D051379:Mice; D010641:Phenotype; D012756:Sheep; D013861:Thiocyanates
D005452:Fluoresceins; D008081:Liposomes; D013861:Thiocyanates; C024098:6-carboxyfluorescein; D016650:Fluorescein-5-isothiocyanate