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Development and validation of streptavidin-biotin-based double antibody sandwich ELISA for ricin diagnosis.

Development and validation of streptavidin-biotin-based double antibody sandwich ELISA for ricin diagnosis.

Authors :
Dixit, Shivani
Parashar, Jagrati
Dhaked, Ram Kumar
Kumar, Abdhesh
Saxena, Nandita
Source :
International Immunopharmacology. May2024, Vol. 132, pN.PAG-N.PAG. 1p.
Publication Year :
2024

Abstract

[Display omitted] • Ricin is a highly toxic ribosome- inactivating protein (RIP) extracted from castor seeds. • It is classified as a scheduled agent by the Centers for Disease Control and Prevention (CDC) and the Chemical Weapons Convention (CWC). • A polyclonal and monoclonal antibody-based sandwich ELISA has been developed with the LOD of 0.45 ng/ml with a linear range between 0.90 and 62.50 ng/ml. • The ELISA is not cross-reactive with similar RIP toxins. • The ELISA can detect ricin in spiked plasma samples and environmental samples. Ricin is a potential biowarfare agent. It is a phytotoxin isolated from castor seeds. At present there is no antidote available for ricin poisoning, patients only get supportive treatment based on their symptoms. This highlights the importance of early detection to avoid severity of accidents and reduce the risk factor. Considering this, our study aimed to develop a highly sensitive and specific sandwich ELISA for the detection of ricin. Ricin was purified from castor seeds. Anti-ricin polyclonal and monoclonal antibodies were generated from rabbit antisera and hybridoma cell (1H6F1) supernatant using a protein A/G column. Antibody titer estimation was done using Indirect ELISA. A streptavidin–biotin-based sandwich ELISA was developed and the limit of detection (LOD), linear range, intra and inter-assay coefficient of variation (CV), and cross-reactivity with other similar toxins were determined. Interference of human plasma samples spiked with ricin was also checked. The LOD of the ELISA was found to be 0.45 ng/ml, with a linear range of 0.90–62 ng/ml, intra and inter-assay CV ranged from 3.34 % to 5 % and 5.17 % to 10.80 % respectively. The assay was not cross-reactive with other similar ribosome-inactivating protein (RIP) toxins. Ricin was detected in spiked plasma samples. The developed assay is highly sensitive and specific for detecting ricin and is not cross-reactive with other similar types of toxins. The assay can detect ricin in spiked plasma samples, so it has the potential to be used for the analysis of clinical samples after ricin poisoning. [ABSTRACT FROM AUTHOR]

Details

Language :
English
ISSN :
15675769
Volume :
132
Database :
Academic Search Index
Journal :
International Immunopharmacology
Publication Type :
Academic Journal
Accession number :
176811579
Full Text :
https://doi.org/10.1016/j.intimp.2024.111986