Characterization of new rat anti-mouse IgE monoclonals and their use along with chimeric IgE to further define the site that interacts with Fc epsilon RII and Fc epsilon RI.
Three rat monoclonal antibodies specific for mouse IgE (C12B9, 23G3, and B1E3) were established by using monoclonal anti-DNP mouse IgE (mIgE) as immunogen. These antibodies, as well as a fourth, (R1E4) were characterized. It was found that one antibody (C12B9) recognizes an allotypic determinant (Igh-7a) found on the C epsilon chain of mIgE. Antibody cross-blocking studies and epitope mapping studies using recombinant mIgE indicated that 3 antibodies (C12B9, R1E4 and 23G3) were directed against the C epsilon 3 domain while one (B1E3) was directed against the C epsilon 4 domain. A highly specific sandwich RIA for mIgE was developed using these antibodies. Use of these monoclonal anti-mIgE antibodies in conjunction with recombinant chimeric mIgE-human IgG1 molecules, demonstrated that the C epsilon 3 domain is important in the binding of mIgE to the murine B cell Fc epsilon RII as well as to the murine mast cell F epsilon RI. The presence of the C epsilon 4 domain influenced the binding of the recombinant IgE to the Fc epsilon RII; in contrast to the C epsilon 4 domain had no effect on binding to the Fc epsilon RI.
Keegan
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AD
Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, MD 21239.
Fratazzi
C
C
Shopes
B
B
Baird
B
B
Conrad
D H
DH
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000944:Antigens, Differentiation, B-Lymphocyte / Q000737:chemistry* / Q000378:metabolism; D001402:B-Lymphocytes / Q000276:immunology*; D001665:Binding Sites; D000939:Epitopes; D006801:Humans; D007073:Immunoglobulin E / Q000737:chemistry / Q000276:immunology*; D007074:Immunoglobulin G / Q000276:immunology; D051379:Mice; D011863:Radioimmunoassay; D051381:Rats; D011961:Receptors, Fc / Q000737:chemistry* / Q000378:metabolism; D017455:Receptors, IgE; D011993:Recombinant Fusion Proteins
D000911:Antibodies, Monoclonal; D000944:Antigens, Differentiation, B-Lymphocyte; D000939:Epitopes; D007074:Immunoglobulin G; D011961:Receptors, Fc; D017455:Receptors, IgE; D011993:Recombinant Fusion Proteins; D007073:Immunoglobulin E