Abstract: The present disclosure provides the complete primary amino acid, and underlying DNA, sequence for the 47-kilodalton membrane immunogen of Treponema pallidum, subsp. pallidum. The sequence was obtained by using a combined strategy of DNA sequencing of the cloned gene as well as confirmatory N-terminal amino acid sequencing of the native antigen. An open reading frame corresponding to the 47-kDa antigen was comprised of 434 amino acid codons. This open reading frame contained a typical 19 amino acid hydrophobic leader peptide flanked by a consensus sequence of Val-Val-Gly-Cys for signal peptidase II, the lipoprotein-specific signal peptidase of prokaryotes. The molecular weight of the mature molecule, excluding modification, is 45,756. Also disclosed are methods for preparing variant and mutant molecules having biologically similar attributes, as well as methods for preparing particular antigenic/immunogenic subportions of the 47-kDa protein.

Abstract: The present disclosure provides the complete primary amino acid, and underlying DNA, sequence for the 47-kilodalton surface immunogen of Treponema pallidum, subsp. pallidum. The sequence was obtained by using a combined strategy of DNA sequencing of the cloned gene as well as confirmatory N-terminal amino acid sequencing of the native antigen. An open reading frame corresponding to the 47-kDa antigen was comprised of 367 amino acid codohs, which gave rise to a calculated molecular weight for the corresponding antigen of about 40,701. Also disclosed are methods for preparing variant and mutant molecules having biologically similar attributes, as well as methods for preparing particular antigenic/immunogenic subportions of the 47-kDa protein. In particular aspects, antigenic/immunogenic subportions are identified by hydrophilicity analysis of the protein sequence.

Abstract: Monoclonal antibodies directed against the 47 kDa major outer membrane surface immunogen of virulent Treponema pallidum were used to select E. coli recombinant clones expressing the 47 kDa immunogen. The phenotype of the clones was dependent on the presence of recombinant plasmid in the host cell. Southern hybridization revealed thatThe United States government may have rights in the substance of this patent because of developmental work supported by the U.S. Department of Health and Human Services in the form of research grants 1-R01-AI-16692 and 1-R01-AI-17366 from NIH-NIAID.

Abstract: Murine anti-Treponema pallidum monoclonal antibodies were employed in the detection of low numbers of pathogenic treponemes. Monoclonal antibodies were used as a primary antibody source in a solid-phase immunoblot assay system. All monoclonal antibodies assayed were capable of detecting ca. 1.0.times.10.sup.3 to 2.5.times.10.sup.3 treponemes. Of 13 monoclonal antibodies examined, 3 were able to detect 10.sup.3 virulent treponemes, and 1 of these antibodies was able to reveal the presence of as few as 500 organisms. Western blot analyses showed that all anti-T. pallidum monoclonal antibodies exhibiting high sensitivities for the detection of T. pallidum cells were directed against an abundant, 47,000-48,000 dalton surface-exposed antigen of the organism. With two possible exceptions, the monoclonal antibodies tested reacted specifically with T.

Abstract: Continuous hybrid cell lines for producing monoclonal antibodies directed against antigens of Treponema pallidum have been developed. The hybrid cell lines were established by fusing differentiated lymphoid cells primed with antigens of Treponema pallidum with hybridoma cells. The resulting fused cells were cloned, isolated, cultured and characterized as to antibody specificity against antigenic determinants of Treponema pallidum.