Selection of antibody probes to correlate protein sequence domains with their structural distribution

We propose a new approach that permits correlation of specific domains defi ned by their primary sequence with their location in the structure of compl ex macromolecular aggregates. It is based on the combination of well-establ ished structural analysis methods that incorporate the use of overlapping p eptides on cellulose membranes for the isolation and purification of specif ic antibodies from a polyclonal antiserum. Monospecific antibodies to the c onnector protein of bacteriophage phi 29 were isolated from polyclonal anti sera using a new development of the spotscan method. These antibodies can b e purified in quantities that allow antigenicity testing in enzyme-linked i mmunosorbent assays, Western blotting and immunoprecipitations, demonstrati ng the specificity of this isolation procedure. This approach has allowed u s to generate direct antibody probes for immunoelectron microscopy mapping of different connector protein domains in a low resolution three-dimensiona l epitope map.