Measurement of Clostridium perfringens beta-toxin production by surface plasmon resonance immunoassay.
A rapid and sensitive assay using surface plasmon resonance (SPR) immunoassay has been developed for the detection of Clostridium perfringens beta-toxin. The SPR immunoassay was conducted off-line by passing fermentation broth by a sensor chip coated with a monoclonal antibody specific for C. perfringens beta-toxin. Quantitation of toxin using SPR immunoassay was achieved by mass transport analysis; results were obtained within 20 min. The SPR immunoassay was compared with an ELISA and the traditional bioassay for C. perfringens beta-toxin. The SPR immunoassay and ELISA detected at least twofold differences in toxin levels at 95% confidence over a broad range of toxin concentrations. The traditional bioassay did not produce the resolution observed with the immunoassays. The SPR immunoassay allows for real-time monitoring of beta-toxin accumulation during production and permits the bioengineer to harvest C. perfringens fermentations when toxin is most concentrated. The SPR methodology may be applied to other fermentations to enhance and optimize toxin yields.
Becton Dickinson Technologies, Research Triangle Park, NC 27709, USA.
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Hsieh
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B
Stewart
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P
Hauer
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P
Haaland
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Campbell
D000818:Animals; D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D001427:Bacterial Toxins / Q000032:analysis* / Q000096:biosynthesis* / Q000276:immunology; D001428:Bacterial Vaccines / Q000096:biosynthesis; D001681:Biological Assay / Q000706:statistics & numerical data; D003016:Clostridium perfringens / Q000276:immunology* / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay / Q000706:statistics & numerical data; D005069:Evaluation Studies as Topic; D007118:Immunoassay / Q000379:methods* / Q000706:statistics & numerical data; D051379:Mice; D012680:Sensitivity and Specificity
D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D001427:Bacterial Toxins; D001428:Bacterial Vaccines; C031884:CPB protein, Clostridium perfringens