Extraction of BoNT/A, /B, /E, and /F with a single, high affinity monoclonal antibody for detection of botulinum neurotoxin by Endopep-MS

• Published in: PloS one Vol. 5; no. 8; p. e12237
• Main Authors: Kalb, Suzanne R, Garcia-Rodriguez, Consuelo, Lou, Jianlong, Baudys, Jakub, Smith, Theresa J, Marks, James D, Smith, Leonard A, Pirkle, James L, Barr, John R
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 17.08.2010; Public Library of Science (PLoS)
• Subjects: Amino Acid Sequence; Animals; Antibodies, Monoclonal - immunology; Antibody Specificity; Antitoxins; Bioassays; Biochemistry; Biochemistry/Protein Chemistry; Biological assays; Biological products; Botulinum toxin; Botulinum Toxins - analysis; Botulinum Toxins - chemistry; Botulinum Toxins - immunology; Botulinum Toxins - isolation & purification; Botulinum Toxins, Type A - analysis; Botulinum Toxins, Type A - chemistry; Botulinum Toxins, Type A - immunology; Botulinum Toxins, Type A - isolation & purification; Botulism; Clostridium botulinum; Clostridium butyricum; Cross Reactions; Disease control; Disease prevention; Endopeptidase; Endopeptidases; Environmental health; Evolution; Female; Food contamination; Gene loci; Genomics; Humans; Immunoglobulins; Immunology; Infectious diseases; Intoxication; Laboratory animals; Limit of Detection; Mass spectrometry; Mass Spectrometry - methods; Mass spectroscopy; Medical research; Mice; Microscopy; Molecular Sequence Data; Monoclonal antibodies; Neurotoxins; Occupational health; Peptides; Pharmaceuticals; Proteins; Public health; Respiratory failure; Serotypes; Tetanus; Toxicology; Toxins; Water-borne diseases; Waterborne diseases; Atlanta Georgia; San Francisco California; Georgia; California; United States > US; Maryland
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0012237

Botulinum neurotoxins (BoNTs) are extremely potent toxins that are capable of causing respiratory failure leading to long-term intensive care or death. The best treatment for botulism includes serotype-specific antitoxins, which are most effective when administered early in the course of the intoxication. Early confirmation of human exposure to any serotype of BoNT is an important public health goal. In previous work, we focused on developing Endopep-MS, a mass spectrometry-based endopeptidase method for detecting and differentiating the seven serotypes (BoNT/A-G) in buffer and BoNT/A, /B, /E, and /F (the four serotypes that commonly affect humans) in clinical samples. We have previously reported the success of antibody-capture to purify and concentrate BoNTs from complex matrices, such as clinical samples. However, to check for any one of the four serotypes of BoNT/A, /B, /E, or /F, each sample is split into 4 aliquots, and tested for the specific serotypes separately. The discovery of a unique monoclonal antibody that recognizes all four serotypes of BoNT/A, /B, /E and /F allows us to perform simultaneous detection of all of them. When applied in conjunction with the Endopep-MS assay, the detection limit for each serotype of BoNT with this multi-specific monoclonal antibody is similar to that obtained when using other serotype-specific antibodies.