Characterization of rat basophilic leukemia cell surface proteins using monoclonal antibodies

Thesis/Dissertation · Fri Jan 01 04:00:00 EST 1988

OSTI ID:5565891

Buonocore-Buzzelli, L M

Rat basophilic leukemia (RBL) cells express both immunoglobulin E (IgE) and immunoglobulin G (IgG) receptors. In this study, mouse monoclonal antibodies were produced against the RBL cell and screened for their ability to precipitate specific bands from {sup 125}I surface labeled cells. Fourteen hybridomas were selected and divided into five groups since many of the hybridomas precipitated bands of identical molecular weight. One or more of the hybridomas from each group, and the cell surface antigens they identified, were further characterized. Binding of all the monoclonal antibodies to the RBL-2H3 cell surface was saturable and of high affinity. In cross inhibition studies, two of the antibodies were found to bind to identical or neighboring epitopes, presumably on the same cell surface molecule. Binding studies using other cell populations demonstrated that the monoclonal antibodies react not only with commonly expressed rat cell surface molecules but also with molecules specifically expressed on rat mast cells and basophils. None of the antibodies were found to induce or inhibit serotonin release from the RBL cells. Western blotting showed most of the antibodies to react with bands whose molecular weights resembled those seen by immuno-precipitation. Antibodies 8 and 12, although from the same group, were found to react with different subunits of the same cell surface protein. Sequential immunoprecipitation and peptide mapping confirmed that the antigens defined by these antibodies were structurally related.

• Research Organization: Catholic Univ. of America, Washington, DC (USA)
• OSTI ID: 5565891
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ANIMAL CELLS
ANIMALS
ANTIBODIES
BIOASSAY
BIOCHEMICAL REACTION KINETICS
CHEMICAL COMPOSITION
DISEASES
ELEMENTS
GLOBULINS
HALOGENS
HEMIC DISEASES
HYBRIDOMAS
IMMUNE SYSTEM DISEASES
IMMUNOASSAY
IMMUNOGLOBULINS
IODINE
ISOTOPE APPLICATIONS
KINETICS
LEUKEMIA
MAMMALS
MEMBRANE PROTEINS
MONOCLONAL ANTIBODIES
NEOPLASMS
NONMETALS
ORGANIC COMPOUNDS
PROTEINS
RATS
REACTION KINETICS
RECEPTORS
RODENTS
TRACER TECHNIQUES
TUMOR CELLS
VERTEBRATES