A Monoclonal Antibody That Recognizes a Common Carbohydrate Epitope Shared by Various Glycoproteins in Human Secretions

The Bovine Lactophorin C-Terminal Fragment and PAS6/7 Were Both Potent in the Inhibition of Human Rotavirus Replication in Cultured Epithelial Cells and the Prevention of Experimental Gastroenteritis

“…1). Lactoferrin from bovine milk 12) exhibited only weak inhibitory activity, with a MIC of 230 mg/ml (Fig. 1).…”

“…12) Rabbit polyclonal antiserum specific for bovine milk fat globule membrane (MFGM) protein PAS6/7 was prepared by Meiji Dairies (Kanagawa, Japan). Mouse monoclonal antibody specific for bovine lactophorin (1C10) was prepared by immunizing mice with bovine MFGM.…”

IEC-6 Intestinal Cell Death Induced by Bovine Milk α-Lactalbumin

“…SDS-PAGE was performed using 15% polyacrylamide precast gels on the mini PROTEAN II electrophoresis cell (Bio-Rad, Hercules, CA) by the method of Laemmli. 9,10) Proteins were dissolved in the sample buffer (0.01 M Tris-HCl buffer pH 6.8 containing 20% glycine, 1% SDS, and 5 mM EDTA, plus 5% -mercaptoethanol) by heating for 5 min in boiling water. Samples (10 ml) were then loaded onto the gel, which was run in a Tris glycine buffer, pH 8.3, with 0.1% SDS.…”

“…Proteins separated by SDS-PAGE were transferred on to a polyvinylidene difluoride (PVDF; Millipore, Bedford, MA) membrane by electrophoresis with a mini Trans-blot electrophoretic transfer cell (Bio-Rad) according to Towbin et al 10,11) For immunoassay, the membrane was rinsed two times with Tris-buffered saline (TBS; 20 mM Tris-HCl, pH 7.5 containing 150 mM NaCl), and incubated at 37…”

Non-Reducing Terminal Fucose within N-Linked Glycan Plays a Significant Role in the Recognition of Human Milk Lactoferrin by the 1CF11 Monoclonal Antibody

“…[7][8][9] We have recently revealed that both hLf and its tryptic digests, after being deglycosylated with N-glycosidase F, showed a significant reduction in their immunoreactivity, although the reaction was never lost. 10) This result clearly indicated that hLf carried two distinct structural elements, the N-glycan-relevant and the N-glycanirrelevant portions, that were recognized by 1CF11 in the molecule.…”