Production of polyclonal and monoclonal antibodies for specific detection of nitrosation-proficient denitrifying bacteria in biological fluids

• Published in: Biochemical and biophysical research communications Vol. 176; no. 1; pp. 262 - 268
• Main Authors: Venezia, N.Dalla, Calmels, S., Bartsch, H.
• Format: Journal Article
• Language: English
• Published: San Diego, CA Elsevier Inc 15.04.1991; Elsevier
• Subjects: Animals; Antibodies; Antibodies, Monoclonal; Bacteria - enzymology; Bacteria - isolation & purification; Bacteriological methods and techniques used in bacteriology; Bacteriology; Biological and medical sciences; Body Fluids - microbiology; cancer; Enzyme-Linked Immunosorbent Assay; enzymes; Female; Fundamental and applied biological sciences. Psychology; Humans; Immunoblotting; Mice; Mice, Inbred BALB C - immunology; Microbiology; monoclonal antibodies; Oxidoreductases - analysis; Oxidoreductases - immunology; Pseudomonas aeruginosa - enzymology; Pseudomonas aeruginosa - isolation & purification; Rabbits - immunology; stomach; urinary bladder; Urine - microbiology; PBS; BSA; SDS; DMSO; Ig; ELISA; Pseudomonadales; Human; Clinical test; Urine; Antibody; Enzyme; Monoclonal antibody; Nitrosation; Immunoblotting assay; Bacteria; Pseudomonadaceae; Pseudomonas aeruginosa; Detection; ELISA assay
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1016/0006-291X(91)90918-W

Polyclonal and monoclonal antibodies were raised against the nitrosating enzyme previously isolated and purified from a denitrifying bacteria Pseudomonas aeruginosa . Using the polyclonal antibodies, a preliminary ELISA test was set up which allowed the detection of the nitrosating enzyme in 2 ml urine samples with a minimal total bacteria count of ≥ 10 5 cells/ml. The use of such a rapid immunological screening test in clinical settings should ascertain whether individual subjects at higher risk for cancer of the stomach or bladder harbour more nitrosation-proficient microorganisms in their microflora. The availability of monoclonal antibodies provides a tool for studying the mechanisms of bacteria-mediated nitrosamine formation from precursor amines.