Capture ELISA showing expression of A, B, H, and Galα1-3Gal antigens on measles.
Microwell plates were coated with a monoclonal antimeasles hemagglutinin Ab. Virus-containing supernatants from cells transfected with either A, B, inactive “O-transferase,” or α1-3GT cDNA clones were diluted with respective supernatant from uninfected cells and added at an identical 5 × 105 PFU/mL. Subsequently, the following detection reagents were added: (A) HRP-labeled A-specific lectin from H pomatia; (B) A-specific mAb BG-2 and an HRP-labeled secondary Ab; (C) B-specific mAb 3E7 and an HRP-labeled secondary Ab; (D) H- or O-specific lectin from U europaeus; (E) an HRP-labeled Galα1-3Gal–specific lectin from B simplicifolia. Plates were subsequently incubated with peroxidase substrate and the absorbance analyzed on a plate reader. Open squares indicate A virus; open circles, B virus; open triangles, H virus; solid squares, Gal(+) virus; solid triangles, Gal(−) virus; and crosses, virus-free supernatant from the respective uninfected cells used to produce virus. In addition, in (A), a filled diamond indicates A virus not incubated with lectin; a filled circle, wells without capture Ab; and a horizontal line, virus which had been preincubated with 10 μg/mL capture Ab. The plotted results indicate the mean (SEM) of duplicate samples.
