Alere Scarborough, Inc.

Chemogen, Inc.

Binax, Inc.

Vladimir Andrei Koulchin

Elena Valentin Molokova

Norman James Moore

Jill L Kerrick

Mary K Konstantin

Mary Kathleen Fent

The present invention involves extracting frombacteria, particularlybacteria, an essentially protein-free O-polysaccharide or carbohydrate antigen, coupling this antigen to an activated chromatographic column through a protein space molecule which is first conjugated to the antigen, utilizing the column thus prepared for the affinity purification of raw polyvalent antibodies to the samebacterium from which the O-polysaccharide or carbohydrate antigen was separated&#x2014;thereby obtaining antigen-specific antibodies which are useful for the rapid detection of the correspondingbacterium or its antibody in human bodily fluids such as urine, sputum, blood and the like or in environmental samples suspected of harboring thebacterium.

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Disclosed is a cell wall C-polysaccharide antigen ofwhich contains not more than 10% by weight of protein, and preferably less which has been purified with 0.1N Na OH prior to deproteinizing. Also disclosed are polyvalent antibodies raised againstwhich have been affinity purified by passing them over a chromatographic affinity matrix to which is coupled the purified and at least partially deproteinized antigen to render them antigen-specific.

The present invention involves extracting frombacteria, particularlybacteria, an essentially protein-free O-polysaccharide or carbohydrate antigen, coupling this antigen to an activated chromatographic column through a protein spacer molecule which is first conjugated to the antigen, utilizing the column thus prepared for the affinity purification of raw polyvalent antibodies to the samebacterium from which the O-polysaccharide or carbohydrate antigen was separated&#x2014;thereby obtaining antigen-specific antibodies which are useful for the rapid detection of the correspondingbacterium or its antibody in human bodily fluids such as urine, sputum, blood and the like or in environmental samples suspected of harboring thebacterium.

The disclosed technology provides an enriched antibody population, highly specific for an antigen of a surface polysaccharide, from a mycobacterium. In a related embodiment, the antibody is enriched by having been raised in an environment that maintains antigenically active antigen. These antibodies may be used in an immunoreactive environment for detecting the presence of a mycobacterial infection in a sample from a subject.

Method of preparing enriched antibodies for detecting mycobacterial infection

Enriched antibody for detecting mycobacterial infection, methods of use and diagnostic test employing same

A process is disclosed for obtaining a C-polysaccharide cell wall antigen containing not more than about 10% protein from bacteria. The antigen thus obtained is conjugated to a spacer molecule, and the free end of the latter is then conjugated to a chromatographic affinity column. The column is then utilized to purify raw antibodies to bacteria, thereby producing antigen-specific antibodies. A portion of such antibodies is conjugated to a labeling agent which displays a visible color change upon reaction of the antibodies with their antigenic binding partner and embedded in a first zone of an immunochromatographic assay device. Another portion of such antibodies is bound to the reaction zone of the device which has a view window. When a liquid sample, such as patient urine, cerebrospinal fluid or blood is applied to the first zone, the conjugate of antibodies and labeling agent and the sample move along a flow strip of bibulous material to the reaction zone wherein, if the sample contains or its cell wall antigen, a sandwich is formed among the labeled conjugate, the antigen and the bound antibodies and a color change is observed. The immunochromatographic assay thus performed is completed within about 15 minutes. This assay affords a basis for rapid and reliable diagnosis of various pathogenic states caused by including pneumonia, bronchitis, otitis media, sinusitis, meningitis, and secondary disease states that commonly occur when primary pneumonic infection caused by this bacterium persists undiminished over a time period of 3-5 days.

Growing community of inventors

Portland, ME, United States of America

Vladimir Andrei Koulchin