Development of ELISA and colloidalgoldimmunoassay for tetrodotoxindetetcionbased on monoclonalantibody

  发布时间: 2016-03-25   信息员:   

Development of ELISA and colloidalgoldimmunoassay for tetrodotoxindetetcionbased on monoclonalantibody

Abstract

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 monoclonalantibody (McAb) with high affinity (about 2.55 × 10(9)) is specific to TTX, and this McAb belongs to the immunoglobulin G (IgG) isotype. Finally, an enzyme-linked immunosorbent assay (ELISA) and colloidalgoldimmunoassay were establishedbased on this McAb. The linear range of ELISA to detect TTX was 5-500 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 colloidalgold 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.

Citation

Ling S, Chen QA, Zhang Y, Wang R, Jin N, Pang J, Wang S (2015) Development of ELISA and colloidal gold immunoassay for tetrodotoxin detetcion based on monoclonal antibody. Biosensors & bioelectronics 71:256-260.



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