title: Extraction and inhibition of enzymatic activity of botulinum neurotoxins /B1, /B2, /B3, /B4, and /B5 by a panel of monoclonal anti-BoNT/B antibodies.
abstract: BACKGROUND
  Botulism is caused by botulinum neurotoxins (BoNTs), extremely toxic proteins which can induce respiratory failure leading to long-term intensive care or death. Treatment for botulism includes administration of antitoxins, which must be administered early in the course of the intoxication; therefore, rapid determination of human exposure to BoNT is an important public health goal. In previous work, our laboratory reported on Endopep-MS, a mass spectrometry-based activity method for detecting and differentiating BoNT/A, /B, /E, and /F in clinical samples. We also demonstrated that antibody-capture is effective for purification and concentration of BoNTs from complex matrices such as clinical samples. However, some antibodies inhibit or neutralize the enzymatic activity of BoNT, so the choice of antibody for toxin extraction is critical.
  
  
  RESULTS
  In this work, we evaluated 24 anti-BoNT/B monoclonal antibodies (mAbs) for their ability to inhibit the in vitro activity of BoNT/B1, /B2, /B3, /B4, and /B5 and to extract those toxins. Among the mAbs, there were significant differences in ability to extract BoNT/B subtypes and inhibitory effect on BoNT catalytic activity. Some of the mAbs tested enhanced the in vitro light chain activity of BoNT/B, suggesting that BoNT/B may undergo conformational change upon binding some mAbs.
  
  
  CONCLUSIONS
  In addition to determining in vitro inhibition abilities of a panel of mAbs against BoNT/B1-/B5, this work has determined B12.2 and 2B18.2 to be the best mAbs for sample preparation before Endopep-MS. These mAb characterizations also have the potential to assist with mechanistic studies of BoNT/B protection and treatment, which is important for studying alternative therapeutics for botulism.
authors.affiliation: Centers for Disease Control and Prevention, National Center for Environmental Health, Division of Laboratory Sciences, 4770 Buford Hwy, N,E,, Atlanta, GA 30341, USA.
authors.forename: Suzanne R
authors.initials: SR
authors.lastname: Kalb
authors.forename: Wanda I
authors.initials: WI
authors.lastname: Santana
authors.forename: Isin N
authors.initials: IN
authors.lastname: Geren
authors.forename: Consuelo
authors.initials: C
authors.lastname: Garcia-Rodriguez
authors.forename: Jianlong
authors.initials: J
authors.lastname: Lou
authors.forename: Theresa J
authors.initials: TJ
authors.lastname: Smith
authors.forename: James D
authors.initials: JD
authors.lastname: Marks
authors.forename: Leonard A
authors.initials: LA
authors.lastname: Smith
authors.forename: James L
authors.initials: JL
authors.lastname: Pirkle
authors.forename: John R
authors.initials: JR
authors.lastname: Barr
mesh_terms: D000911:Antibodies, Monoclonal / Q000494:pharmacology*; D001905:Botulinum Toxins / Q000037:antagonists & inhibitors* / Q000276:immunology / Q000302:isolation & purification*; D019274:Botulinum Toxins, Type A; D001906:Botulism / Q000235:genetics / Q000378:metabolism*; D003014:Clostridium botulinum / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism; D018604:Epitope Mapping; D006801:Humans
chemical_list: D000911:Antibodies, Monoclonal; C096323:rimabotulinumtoxinB; D001905:Botulinum Toxins; D019274:Botulinum Toxins, Type A
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