Journal
BIOSENSORS & BIOELECTRONICS
Volume 71, Issue -, Pages 256-260Publisher
ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2015.04.049
Keywords
Tetrodotoxin; ELISA; Monoclonal antibody; Colloidal gold immunoassay
Categories
Funding
- Natural Science Foundation of Fujian Province [2011J05049]
- Key Scientific and Technology Project of Fujian Province of China [2014YZ0105, 2012Y0002]
- State Key Laboratory of Freshwater Ecology and Biotechnology [2009FB13]
- Fujian Development and Reform Commission
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A monoclonal hybridoma cell named 5B9 against tetrodotoxin (TTX) was obtained after fusion of myeloma SP2/0 cells with spleen cells isolated from the immunized Balb/c mice. The 5B9 monoclonal antibody (McAb) with high affinity (about 2.55 x 10) is specific to TTX, and this McAb belongs to the immunoglobulin G (IgG) isotype. Finally, an enzyme-linked immunosorbent assay (ELISA) and colloidal gold immunoassay were established based on this McAb. The linear range of ELISA to detect TTX was 5500 ng/mL, and the limit of detection (LOD) was 4.44 ng/mL. The average CV of intra- and inter-assay was less than 8%, with the samples recovery range of 70.93-99.99%. A competitive format colloidal gold strip was developed for detection of TTX in real samples, and the LOD for TTX is 20 ng/mL, and the assay time of the qualitative test can be finished in less than 10 min without any equipment. The result from test strip revealed that the test strip has a good agreement with those obtained from ELISA. (C) 2015 Elsevier B.V. All rights reserved.
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