Production and characterization of monoclonal antibodies directed against bovine milk fat globule membrane (MFGM).
Nine hybridomas secreting monoclonal antibodies (mAbs) to bovine milk fat globule membrane (MFGM) were produced from spleen cells of three immunized BALB/c mice. Several MFGM antigens recognized by some mAbs were identified as a 120 kDa protein and 67 kDa (butyrophilin), 57 kDa (PAS-6), 53 kDa (PAS-7), 33 kDa glycoproteins. The other mAbs secreted by four independent hybridoma clones recognized many broad bands ranging from 20 to 200 kDa. The 120 kDa protein and 67 kDa, 57 kDa, 53 kDa glycoproteins were detected by each mAb in the plasma membrane fraction prepared from a lactating bovine mammary gland. Moreover, mammary gland epithelium of a thin section was specifically stained with these mAbs, indicating that these mAbs directed against MFGM recognized membrane proteins and glycoproteins of lactating mammary epithelial cells. Upon heating of the MFGM in phosphate buffer, pH 7.4 at 100 degrees C for 10 min, the antigens still retained most of its reactivity to these mAbs, whereas, proteolytic cleavage by trypsin and chymotrypsin strongly reduced its reactivity to these mAbs by 60% or more except for two mAbs which recognized the 57 and 53 kDa glycoproteins, respectively.
Aoki
N
N
Department of Applied Biological Sciences, School of Agricultural Sciences, Nagoya University, Japan.
Kuroda
H
H
Urabe
M
M
Taniguchi
Y
Y
Adachi
T
T
Nakamura
R
R
Matsuda
T
T
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D001940:Breast / Q000737:chemistry; D002417:Cattle; D002462:Cell Membrane / Q000737:chemistry; D010450:Endopeptidases; D006358:Hot Temperature; D007150:Immunohistochemistry; D007774:Lactation; D008562:Membrane Glycoproteins / Q000032:analysis / Q000276:immunology*; D008565:Membrane Proteins / Q000032:analysis / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D018396:Mucin-1
D000911:Antibodies, Monoclonal; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D018396:Mucin-1; D010450:Endopeptidases