Purified antigen to test for Neisseria gonorrhoeae antibodies

Claim 1

A heat stabile antigen preparation, protein in nature isolated from Neisseria gonorrhoeae and useful for detecting the presence of Neisseria gonorrhoeae antibodies in human serum, containing a heat stable, species specific L-antigen which is characterized as follows: 1. stable in boiling water for a period of one hour, 2. stable in an aqueous medium at pH values of 3-11, 3. stable when incubated with the following enzymes: DNA-ase RNA-ase dextranase neuraminidase lysosyme,

Claim 10

A method for the preparation of an antigen preparation protein in nature and useful for detecting the presence of Neisseria gonorrhoeae antibodies in human serum, containing a heat stable, species specific antigen characterized as follows: 1. stable in boiling water for a period of one hour, 2. stable in an aqueous medium at pH values of 3-11, 3. stable when incubated with the following enzymes: DNA-ase RNA-ase dextranase neuraminidase lysosyme, 4. isoelectric point of 4±0.2, 5 molecular weight of active subunits of 37,000-40,000 as determined by SDS-polyacrylamide gel, 6. contains 2-3% organic phosphorus, 7. soluble in water, and more soluble in aqueous media containing surface active agents; and insoluble in methanol, chloroform and acetone, 8. reacts specifically with antibodies produced in rabbits against purified L-antigen, and 9. partially soluble in 5% to 10% trichloroacetic acids; 10. contains less than 1% carbohydrate, 11. inactivated when incubated with trypsin, which method comprises the steps of: 1. extracting a cell pellet of Neisseria gonorrhoeae with a solution containing a detergent, or an inorganic salt, 2. passing resulting extract over a column containing a molecular sieve with an exclusion limit of at least 200,000, 3. adding trichloroacetic acetic acid at a concentration of from 5% to 10% by volume to precipitate impurities; and 4. separating said impurities from a solution containing said antigen.

Claim 11

A method for the preparation of an antigen preparation protein in nature and useful for detecting the presence of Neisseria gonorrhoeae antibodies in human serum, containing a heat stable, species specific antigen characterized as follows: 1. stable in boiling water for a period of one hour, 2. stable in an aqueous medium at pH values of 3-11, 3. stable when incubated with the following enzymes: DNA-ase RNA-ase dextranase neuraminidase lysosyme, 4. isoelectric point of 4±0.2, 5 molecular weight of active subunits of 37,000-40,000 as determined by SDS-polyacrylamide gel, 6. contains 2-3% organic phosphorus, 7. soluble in water, and more soluble in aqueous media containing surface active agents; and insoluble in methanol, chloroform and acetone, 8. reacts specifically with antibodies produced in rabbits against purified L-antigen, and 9. partially soluble in 5% to 10% trichloroacetic acids; 10. contains less than 1% carbohydrate, 11. inactivated when incubated with trypsin, which method comprises the steps of: 1. extracting a cell pellet of Neisseria gonorrhoeae with a solution containing a detergent, or an inorganic salt, 2. passing resulting extract over a column containing a molecular sieve with an exclusion limit of at least 200,000, 3. adding trichloroacetic acetic acid at a concentration of from 5% to 10% by volume to precipitate impurities; and 4. separating said impurities from a solution containing said antigen.