Homogeneous fluoroimmunoassay using Lucifer yellow VS: determination of albumin plasma

Non-separation fluoroimmunoassays are well suited to automation. The potential sensitivity of such assays has not been realised because the most commonly used label (fluorescein) has absorption and emission spectra which coincide with those of blood constituents and because it has a very small Stokes shift. Lucifer yellow VS has a larger Stokes shift than fluorescein and an emission maximum at longer wavelength. We describe an energy transfer fluoroimmunoassay for plasma albumin in which Lucifer yellow VS is used to label albumin and rhodamine B isothiocyanate is used to label anti-albumin antibodies. The assay shows good correlation with a dye-binding method for albumin and has sensitivity and precision which compare favourably with similar assays using a fluorescein label.