Agglutinating activities of pepsin fragments and subunits of IgM antibodies.
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Biomedical subjects
Publications and source records attributed to K Onoue.
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Immunoglobulin M rabbit antibodies to a hapten are shown to have ten binding sites per molecule. The affinity for the specific hapten is approximately 100 times greater for one-half of the sites than for the other half. All sites are retained in the five 7S subunits produced by reduction and alkylation of the immunoglobulin M. Each of the 7S subunits of the IgM molecule apparently has one strong and one weak site.
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Multiple antibody components of rabbit antisera against p-azobenzenearsonate (R(p)) were studied with respect to their globulin nature and skin-sensitizing activity. IgA antibody was characterized by isolating two IgA-rich fractions from a specifically purified antibody preparation. Examination of these fractions showed that IgA antibodies existed in two molecular forms, one with a sedimentation constant of 7S and the other 9S. Skin-sensitizing activity was examined by a P-K type test and a PCA test with R(p)-rabbit serum albumin in homologous (rabbit) species. Only the 7S but not 9S IgA antibody sensitized rabbit skin. IgM antibody showed no activity and IgG antibody showed very low activity. In contrast, only IgG antibody was active in the P-K type test to sensitize a heterologous species (guinea pig). None of the antibodies of other classes showed sensitizing activity in heterologous skin. The 7S IgA antibody lost its sensitizing activity upon reduction and alkylation, although no change in its molecular size could be observed. The loss of sensitizing activity was not due to the destruction of antigen-binding activity since the treated 7S IgA antibody retained this activity as shown by radioimmunoelectrophoresis and by binding to the specific immunoadsorbent. The 9S IgA antibody was more resistant to these treatments than the IgM antibody and showed no indication of dissociation. The treated 9S IgA also retained antigen-binding activity. Both the P-K type and PCA reactions were considerably stronger when the interval between injections of antibody and antigen was 24 hr rather than 4 to 5 hr.
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