Human anti-IgE antibodies by repertoire cloning.

Abstract

Anti-IgE antibodies have been detected in sera of patients with an allergic disease where they might play a role in the regulation of (Ig)E-mediated reactions. Using a recombinant phage surface display expression system a combinatorial library of antibody heavy and light chains was constructed from peripheral blood mononuclear cells from an atopic donor immunized with tetanus toxoid. Screening of the library allowed the identification of a large number of phages displaying human monovalent antigen-binding fragments (Phab) against tetanus toxoid and IgE. Surprisingly, we found a high frequency of Phab against particular IgE myelomas that was comparable to the frequency found for Phab against tetanus toxoid. However, most of these Phab were directed to different idiotypic determinants, depending on the IgE myeloma used for the panning procedure. Nevertheless, two clones were found to have anti-isotypic specificity and were shown to react specifically with the CH2 domain of the IgE heavy chain.