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Simultaneous and sensitive detection of six serotypes of botulinum neurotoxin using enzyme-linked immunosorbent assay-based protein antibody microarrays.*

Posted by on in 2012
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Zhang YLou JJenko KLMarks JDVarnum SM. 2012. Anal Biochem.  1430(2):185-92. doi: 10.1016/j.ab.2012.08.021. Epub 2012 Aug 27.

Biological Sciences Division, Pacific Northwest National Laboratory, Richland, WA 99352, USA.

Abstract

Botulinum neurotoxins (BoNTs), produced by Clostridium botulinum, are a group of seven (A-G) immunologically distinct proteins and cause the paralytic disease botulism. These toxins are the most poisonous substances known to humans and are potential bioweapon agents. Therefore, it is necessary to develop highly sensitive assays for the detection of BoNTs in both clinical and environmental samples. In the current study, we have developed an enzyme-linked immunosorbent assay (ELISA)-based protein antibody microarray for the sensitive and simultaneous detection of BoNT serotypes A, B, C, D, E, and F. With engineered high-affinity antibodies, the BoNT assays have sensitivities in buffer ranging from 1.3fM (0.2pg/ml) to 14.7fM (2.2pg/ml). Using clinical and food matrices (serum and milk), the microarray is capable of detecting BoNT serotypes A to F to similar levels as in standard buffer. Cross-reactivity between assays for individual serotype was also analyzed. These simultaneous, rapid, and sensitive assays have the potential to measure botulinum toxins in a high-throughput manner in complex clinical, food, and environmental samples.

*The Chicken anti-Botulinum Neurotoxin B used in this publication was manufactured by Gallus Immunotech Inc.

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Copyright © 2012 Elsevier Inc. All rights reserved.

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