Polyethylene glycol-enhanced nephelometric quantification of bovine submaxillary mucin with Helix pomatia lectin.
A new principle for quantification of intact mucin has been developed. The method is reproducible, simple, and sensitive at the submicrogram level and can be performed within an hour. The procedure involves incubation of mucin (bovine submaxillary mucin) and polyethylene glycol with the lectin from the snail Helix pomatia and the complexes formed are quantitated by nephelometry. Polyethylene glycol was found to enhance the affinitin reaction in a dose-dependent fashion. Increasing concentrations of acetate or citrate led to decreasing light scattering of the polyethylene glycol--bovine submaxillary mucin--H. pomatia lectin reaction mixture. Results from displacement studies with N-acetyl-galactosamine suggest that H. pomatia lectin may contain an allosteric type of regulatory site. The quantitative results obtained with bovine submaxillary mucin using the new method correlated well with those obtained with an established method for neuraminic acid determination after neuraminidase digestion (R = 0.969, P less than 0.001; n = 24).
D000818:Animals; D002021:Buffers; D002417:Cattle; D006863:Hydrogen-Ion Concentration; D037102:Lectins; D009077:Mucins / Q000032:analysis*; D009391:Nephelometry and Turbidimetry; D011092:Polyethylene Glycols; D012471:Salivary Proteins and Peptides / Q000032:analysis*; D013363:Submandibular Gland
D002021:Buffers; C038095:Helix lectin; D037102:Lectins; D009077:Mucins; D012471:Salivary Proteins and Peptides; D011092:Polyethylene Glycols