Development of ELISA and colloidal gold immunoassay for tetrodotoxin detetcion based on monoclonal antibody.
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 × 10(9)) 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 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 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.
The Ministry of Education Key Laboratory of Biopesticide and Chemical Biology, Fujian Key Laboratory of Pathogenic Fungi and Mycotoxins, Fujian Agriculture and Forestry University, Fuzhou 350002, China; School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, China.
Sumei
S
Ling
Fujian Commercial College, Fuzhou 350012, China; School of Food Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, China.
Qing-Ai
QA
Chen
The Ministry of Education Key Laboratory of Biopesticide and Chemical Biology, Fujian Key Laboratory of Pathogenic Fungi and Mycotoxins, Fujian Agriculture and Forestry University, Fuzhou 350002, China; School of Food Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, China.
Yuming
Y
Zhang
The Ministry of Education Key Laboratory of Biopesticide and Chemical Biology, Fujian Key Laboratory of Pathogenic Fungi and Mycotoxins, Fujian Agriculture and Forestry University, Fuzhou 350002, China; School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, China.
Rongzhi
R
Wang
The Ministry of Education Key Laboratory of Biopesticide and Chemical Biology, Fujian Key Laboratory of Pathogenic Fungi and Mycotoxins, Fujian Agriculture and Forestry University, Fuzhou 350002, China; School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, China.
Ni
N
Jin
School of Food Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, China.
Jie
J
Pang
The Ministry of Education Key Laboratory of Biopesticide and Chemical Biology, Fujian Key Laboratory of Pathogenic Fungi and Mycotoxins, Fujian Agriculture and Forestry University, Fuzhou 350002, China; School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, China. Electronic address: wshyyl@sinna.com.
Shihua
S
Wang
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D017979:Gold Colloid / Q000737:chemistry*; D006825:Hybridomas / Q000276:immunology; D057230:Limit of Detection; D008387:Marine Toxins / Q000032:analysis* / Q000276:immunology; D008807:Mice, Inbred BALB C; D023721:Tetraodontiformes; D013779:Tetrodotoxin / Q000032:analysis* / Q000276:immunology
D000911:Antibodies, Monoclonal; D017979:Gold Colloid; D008387:Marine Toxins; D013779:Tetrodotoxin
Colloidal gold immunoassay; ELISA; Monoclonal antibody; Tetrodotoxin