Detection of Staphylococcal enterotoxin B via biomolecular interaction analysis mass spectrometry.
Detection of Staphylococcus enterotoxin B (SEB) by biomolecular interaction analysis mass spectrometry (BIA/MS) is presented in this work. The BIA/MS experiments were based on a surface plasmon resonance (SPR) MS immunoassay that detects affinity-captured SEB both via SPR and by means of exact and direct mass measurement by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry. Experiments were performed with standard samples and food samples to assess the BIA/MS limit of detection for SEB and to set the experimental parameters for proper quantitation. Single and double SPR referencing was performed to accurately estimate the amount of the bound toxin. Reproducible detection of 1 ng of SEB per ml, corresponding to affinity capture and MS analysis of approximately 500 amol of SEB, was readily achieved from both the standard and mushroom samples. A certain amount of SEB degradation was indicated by the signals in the mass spectra. The combination of MS with SPR-based methods of detection creates a unique approach capable of quantifying and qualitatively analyzing protein toxins from pathogenic organisms.
Nedelkov
Dobrin
D
Intrinsic Bioprobes Inc., Tempe, Arizona 85281, USA. dnedelkov@intrinsicbio.com
Nelson
Randall W
RW
D000942:Antigens, Bacterial / Q000032:analysis* / Q000737:chemistry* / Q000302:isolation & purification; D004768:Enterotoxins / Q000032:analysis* / Q000737:chemistry* / Q000302:isolation & purification; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D012680:Sensitivity and Specificity; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods; D013210:Staphylococcus
D000942:Antigens, Bacterial; D004768:Enterotoxins; C031818:enterotoxin B, staphylococcal