ABSTRACT
The uniformity of Xenobind covalent binding microwell plates was compared with that of passive binding plates by binding streptavidin to the plate surface, reacting the streptavidin with biotinylated alkaline phosphatase, and developing color with phenolphthalein monophosphate. Absorbance of the covalent binding plates at 570 nm was 0.95 A (CV 1.9%) compared with 0.35-0.5 A (CV 6.7-16.9%), for passive plates. No edge effect was observed with Xenobind covalent plates. Use of both plates in a model immunoassay gave a CV of 4% for the covalent binding plate and 12-15% for the passive binding plates. Moreover, the background absorbance (color development when no antigen was present) for the covalent binding plates was < 0.05 A in all cases, whereas that for the passive binding plates ranged up to 0.45 A. The high background on passive plates results from displacement of capture antibody and (or) blocking agent from the plate, thereby providing a place for detector antibody binding.
Subject(s)
Immunoassay/methods , Absorption , Alkaline Phosphatase , Antibodies, Monoclonal , Bacterial Proteins , Binding Sites, Antibody , Phenolphthaleins , Reproducibility of Results , Streptavidin , ThermodynamicsABSTRACT
The effect of UV-B radiation on the crystalline lens was examined by subjecting bovine lenses in culture to varying low exposures at 300 nm. Lens optical quality was monitored on a long-term basis (to 1000 hrs.) with an automated scanning laser system that recorded both change in relative scatter and focal length across each lens. Data were collected for 20 lens positions at each scan. Radiant exposure levels consisted of 0.5, 0.25, 0.125, 0.06 and 0.03 Jcm-2. Twenty irradiated lenses were compared to twelve untreated controls. All of the irradiated lenses showed changes in scatter and focal length relative to the controls. Most (about 75%) of the treated lenses showed significant increases in scatter (200-400%) and focal length (10-20%) at 40 to 60 hours after exposure. A similar time frame for lens damage was noted by visual inspection. Exposure to UV-B at the above doses did not affect culture longevity.
