A method is presented whereby antigenic determinants recognized by specific monoclonal antibodies can be mapped to specific sites on a protein sequence with high resolution. Short DNase I-generated DNA fragments encoding portions of the protein of interest are molecularly cloned into the EcoRI site of the fi-galactosid.Ise gene of phage A Charon 16 so as to obtain expression of randojn protein fragments as fusion proteins. The monoclonal antibody is used to screen the phage library to isolate phage expressing the specific antigenic determinant. DNA of immunoreactive phage can be analyzed rapidly and subcloned to allow DNA sequence determination.The method is generally applicable and permits antigenic determinants of functionally interesting monoclonal antibodies to be mapped and related to specific protein &equences. We have used this procedure to determine the region of the feline leukemia virus envelope protein gp7Q recognized by a virusneutralizing monoclonal antibody, cl.25. Antibocy binding was mapped to a }4-amino acid region in the amino-terminal half of gp7O. This region way be directly involved in an essential function of the gp7O protein, perhaps in gp7O-mediated host recognition functions. Synthetic peptides derived from this region may provide useful vaccine antigens for the prevention of feline leukemia viru4-ilssociated disease in cats.

We used the protein gel transfer technique (Western blotting) to analyze the immune response to individual electrophoretically resolved Toxoplasma antigens. Toxoplasma antigens were fractionated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The array of resolved proteins was electrophoretically transferred to CNBr-activated filter paper and incubated with serum samples from patients with either acute or chronic Toxoplasma infections or with serum samples from seronegative controls. Serum immunoglobulin G and immunoglobulin M bound to individual Toxoplasma antigens were detected by radioiodinated staphylococcal protein A and a goat anti-human immunoglobulin M antibody, respectively. A complex array of Toxoplasma antigens was revealed by this procedure. Immunoglobulin M directed against a low-molecular-weight antigen was detected in sera from all acutely infected adults (19 sera tested) and was absent from all sera from individuals with chronic infections (9 sera tested) and from sera from seronegative controls (9 sera tested).

A recombinant plasmid containing a 5.6-kilobase-pair DNA fragment of the Treponema pallidum genome was characterized by endonuclease mapping, and the encoded proteins were expressed in Escherichia coli and analyzed by use of in vitro transcription and translation. One of the proteins, identified as having a molecular weight of 37,000 (37K protein), was selected for further study. Initially, the seroreactivity of the partially purified 37K antigen was demonstrated by immunoblotting. After its purification to near homogeneity, the cloned T. pallidum protein was assessed for diagnostic significance by radioimmunoassay. Although first identified as seroreactive by screening with secondary syphilitic sera (T.