C - Chemistry – Metallurgy – 12 – N
Patent
C - Chemistry, Metallurgy
12
N
167/37, 195/1.22
C12N 15/31 (2006.01) C07K 14/20 (2006.01) C12N 1/21 (2006.01) C12N 15/70 (2006.01) C12Q 1/68 (2006.01) G01N 33/571 (2006.01) A61K 39/00 (2006.01)
Patent
CA 1336692
Monoclonal antibodies directed against the 47 kDa major outer membrane surface immunogen of virulent Tre- ponema 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 that the cloned T. pallidum DNA sequence was an accurate representation of the T. pallidum genomic DNA arrangement. Purified IgG from rabbits experimentally infected with T. pallidum and human secondary syphilitic sera specifically reacted with the clones while normal human serum or IgG from normal rabbit serum did not. Results of Southern hybridization indicated that a homologous 47 kDa immunogen gene was absent in at least 4 species of nonpathogenic treponemes tested, as well as from total rabbit genomic DNA. Rabbit anti-T. phagedenis biotype Reiter (treponemal nonpathogen) antiserum and a monoclonal antibody directed against a "common" treponemal determinant were unreactive with the clones. Western blotting and radioimmunoprecipi- tation experiments with specific monoclonal antibodies revealed that the recombinant (E. coli) and native (T. pallidum) forms of the antigen had identical electro- phoretic mobilities. The availability of recombinant 47 kDa immunogen provides a new opportunity for biochemical analysis of the protein, structure-function studies, examination of its role in microbial pathogenesis, and assessment of its diagnostic and vaccinogenic potentials.
547633
Board Of Regents The University Of Texas System
Riches Mckenzie & Herbert Llp
LandOfFree
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