Figure 5

Neuraminidase treatment enhances 45M1 performance in ELISA. Mucin secretions were collected from GPTE cell cultures on day 9 after exposure to air. Samples were treated with neuraminidase (0.625 mU/μg total protein) for 2 hrs at 37°C [35]. Both neuraminidase-treated and untreated samples were diluted 2-fold in series, then coated on ELISA plates in duplicate. a) Neuraminidase treatment slightly increased absorbance values using C4 antibody (diluted 1:500), although the logarithmic slope and trend line fit of both treated and non-treated samples remained similar. b) Neuraminidase treatment improved the linear range of protein detection in an ELISA using the 45M1 monoclonal antibody (diluted 1:1000), as indicated by better R² value fit (calculated for ranges of increasing absorbance with increasing concentration) of the logarithmic trend line. Specifically, treatment extended the linear detection range above 35 ng/well.