Time-dependent study of the antibody response to sperm-whale myoglobin: recognition of the antigenic sites is unaltered over an extended period of immunization

We have recently shown that the antigenic structure of sperm-whale myoglobin is not dependent on the host species in which the antisera are raised. The purpose of the present work was to determine whether or not the molecular immune recognition of a protein by antibody is subject to a time-dependency. We have investigated the recognition of the antigenic sites by serial antisera obtained in two rabbits at different times after the initial immunization, from the earliest bleeding with detectable antibodies (9 days) up to a year. The specificity of these antisera was determined by their cross-reactions with 13 myoglobins from different species. The reactivity was measured by quantitative immunoadsorbent titration studies from the amount of radioiodinated antibodies that could be bound maximally (i.e. plateau binding values) by immunoadsorbents of each myoglobin. From these studies and our recent structural analysis, which enabled us to identify for each of these myoglobins the regions retaining reactivity with antibodies to sperm-whale myoglobin, we have concluded that following maturation of the immune response the antigenic recognition of a protein (i.e. the regions that are recognized as antigenic) is not dependent upon the time antisera are obtained after the first immunization. In the early periods of the response, fluctuations are observed in the relative immunodominance of the sites. This further confirms our earlier conclusions that the antigenic sites on a protein are structurally inherent in the protein.