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AssigneeResearch Corporation
InventorsHarlan D. Caldwell, Cho-Chou Kuo, George E. Kenny
Estimating the term…
Solubilized antigens of C. trachomatis strain LGV-434 upon analysis using two-dimensional immunoelectrophoresis yielded a single antigen which was found to be consistently precipitated by sera of patients with C. trachomatis infections. This antigen as antigen-antibody complex was employed as an immunogen to prepare a rabbit monospecific antiserum to this component or antigen. This monospecific antiserum demonstrated the presence of the antigen in each of the 15 strains of C. trachomatis organisms and was non-reactive with strains of C. psittaci. The C. trachomatis specific antigen was purified by immunoadsorption chromatography using monospecific antiserum as a specific ligand covalently bound to agarose gel columns and the resulting purified antigen employed to detect antibody from the sera of lymphogranuloma venereum patients using counterimmunoelectrophoresis. When the C. trachomatis specific antigen is isotopically labeled and utilized in the highly sensitive radioimmune assay antibody to the antigen should be demonstrated and a serological test based thereon should be applicable for the serological diagnosis of non-gonococcal urethritis (NGU).
Chlamydia trachomatis specific antigen useful for the serological diagnosis of lymphogranuloma venereum, said antigen having a molecular weight of about 160,000, being stable to ribonuclease and deoxyribonuclease and at a pH in the range 2.2-10.6 but whose antigenicity is destroyed by protease treatment and heating at a temperature of 56.degree. C. for about 30 minutes.
A method of preparing Chlamydia trachomatis antigen which is consistently precipitated by sera of patients with C. trachomatis infections which comprises solubilizing C. trachomatis in the presence of a non-ionic surfactant and recovering the resulting solubilized C. trachomatis antigen by immunoadsorption chromatography employing monospecific antiserum to said antigen as a specific ligand for said antigen.
A method of isolating Chlamydia trachomatis antigen in the form of its antigen-antibody complex which comprises solubilizing C. trachomatis antigen and subjecting the resulting solubilized antigen to two-dimensional immunoelectrophoresis and recovering the antigen-antibody complex.
A method of preparing monospecific antiserum to antigen of Chamydia trachomatis which comprises solubilizing the antigens of C. trachomatis in the presence of a non-ionic surfactant, subjecting the resulting solubilized antigens to two-dimensional immunoelectrophoresis on agarose, thereby precipitating the C. trachomatis antigen as an antigen-antibody complex, recovering said antigen-antibody complex, introducing said resulting recovered antigen-antibody complex into rabbit, thereby causing the production of antiserum therein and recovering rabbit serum containing the resulting produced antiserum.
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