A Novel Colourimetric Homogeneous Liposomal Immunoassay Using Sulphorhodamine B

The dye, Sulphorhodamine B, was entrapped within liposomes prepared by the reverse evaporation technique. Marked differences in absorption spectra were found when free and entrapped dyes were compared, with a shift in the wavelength of maximum absorption. When entrapped dye was released by lysis of the liposomes, for example by detergent, the absorption spectrum reverted to that of free dye. This absorption change was employed in a novel marker system for complement-mediated immunoassay. As a model assay, human serum albumin was measured using this approach. Liposomes which had been coated with albumin were incubated with anti-albumin antibody and complement and the resulting absorption change measured using an automated spectrophotometric analyser. The decrease in absorption change on the addition of albumin formed the basis of a competitive homogeneous immunoassay for human serum albumin. Using purified albumin as a standard, a correlation of 0.96 was obtained when the albumin in human serum was measured in the liposomal assay and the results compared to measurements using a bromocresol green method.