Library of monoclonal antibodies against brush border membrane epithelial antigens
A purified fraction of proximal tubule brush border membranes (BBM) was prepared from dog kidney and used to immunize mice. The standard technique of hybridoma production was followed as described by Kohler and Milstein. Production of antibodies was detected by indirect immunofluorescence on dog kidney slices and by immunodot against the purified fraction on nitrocellulose. Five hybrids exhibited anti BBM activity. These were cloned twice and yielded stable cell lines producing IgG type monoclonal antibodies against BBM. They were designated A/sub 1/, C/sub 7/, D/sub 3/, D/sub 7/ and H/sub 4/. As a family these five monoclonals have broad tissue specificity, i.e. positive staining of the surface mucosa of intestinal kidney proximal tubules. D/sub 3/ exhibits even broader specificity for epithelium reacting with bile canaliculi and choroid plexus. The authors have verified that at least 4/5 antibodies are directed against BBM protein as revealed by immunoprecipitation of solubilized BBM and detected by Coomassie blue staining or autoradiography of lactoperoxidase labelled BBM. Most interestingly all antibodies bind to the surface of LL CPK/sub 1/ cells, a continuous pig kidney cell line of undefined origin but exhibiting many characteristics of proximal tubule cells. The library of monoclonal antibodies obtained provide important probes with which to study membrane biogenesis and polarization in epithelial cells.
- Research Organization:
- Univ. of Toronto, Ontario
- OSTI ID:
- 5385805
- Report Number(s):
- CONF-8604222-; TRN: 86-026565
- Journal Information:
- Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States), Vol. 45:3; Conference: 70. annual meeting of the Federation of American Society for Experimental Biology, St. Louis, MO, USA, 13 Apr 1986
- Country of Publication:
- United States
- Language:
- English
Similar Records
Diagnosis of and therapy for solid tumors with radiolabeled antibodies and immune fragments
Production and characterization of a monoclonal antibody to H+ATPase
Related Subjects
EPITHELIUM
IMMUNE REACTIONS
MONOCLONAL ANTIBODIES
BIOCHEMICAL REACTION KINETICS
ANTIGENS
AUTORADIOGRAPHY
CELL MEMBRANES
DOGS
FLUORESCENCE
HYBRIDOMAS
KIDNEYS
MICE
PURIFICATION
TUBULES
ANIMAL CELLS
ANIMAL TISSUES
ANIMALS
ANTIBODIES
BODY
CELL CONSTITUENTS
KINETICS
LUMINESCENCE
MAMMALS
MEMBRANES
ORGANS
REACTION KINETICS
RODENTS
TISSUES
VERTEBRATES
550201* - Biochemistry- Tracer Techniques