Development of a New Monoclonal Antibody against Brevetoxins in Oyster Samples Based on the Indirect Competitive Enzyme-Linked Immunosorbent Assay
Xiya Zhang,
Mingyue Ding,
Chensi Zhang,
Yexuan Mao,
Youyi Wang,
Peipei Li,
Haiyang Jiang,
Zhanhui Wang,
Xuezhi Yu
Affiliations
Xiya Zhang
Henan Province Engineering Research Center for Food Safety Control of Processing and Circulation, College of Food Science and Technology, Henan Agricultural University, 63 Nongye Road, Zhengzhou 450002, China
Mingyue Ding
Henan Province Engineering Research Center for Food Safety Control of Processing and Circulation, College of Food Science and Technology, Henan Agricultural University, 63 Nongye Road, Zhengzhou 450002, China
Chensi Zhang
College of Life Sciences, Henan Agricultural University, 63 Nongye Road, Zhengzhou 450002, China
Yexuan Mao
Henan Province Engineering Research Center for Food Safety Control of Processing and Circulation, College of Food Science and Technology, Henan Agricultural University, 63 Nongye Road, Zhengzhou 450002, China
Youyi Wang
Henan Province Engineering Research Center for Food Safety Control of Processing and Circulation, College of Food Science and Technology, Henan Agricultural University, 63 Nongye Road, Zhengzhou 450002, China
Peipei Li
Beijing Key Laboratory of Detection Technology for Animal–Derived Food Safety, Beijing Laboratory of Food Quality and Safety, College of Veterinary Medicine, China Agricultural University, Beijing 100193, China
Haiyang Jiang
Beijing Key Laboratory of Detection Technology for Animal–Derived Food Safety, Beijing Laboratory of Food Quality and Safety, College of Veterinary Medicine, China Agricultural University, Beijing 100193, China
Zhanhui Wang
Beijing Key Laboratory of Detection Technology for Animal–Derived Food Safety, Beijing Laboratory of Food Quality and Safety, College of Veterinary Medicine, China Agricultural University, Beijing 100193, China
Xuezhi Yu
Beijing Key Laboratory of Detection Technology for Animal–Derived Food Safety, Beijing Laboratory of Food Quality and Safety, College of Veterinary Medicine, China Agricultural University, Beijing 100193, China
The consumption of shellfish contaminated with brevetoxins, a family of ladder-frame polyether toxins formed during blooms of the marine dinoflagellate Karenia brevis, can cause neurotoxic poisoning, leading to gastroenteritis and neurotoxic effects. To rapidly monitor brevetoxin levels in oysters, we generated a broad-spectrum antibody against brevetoxin 2 (PbTx-2), 1 (PbTx-1), and 3 (PbTx-3) and developed a rapid indirect competitive enzyme-linked immunosorbent assay (icELISA). PbTx-2 was reacted with carboxymethoxylamine hemihydrochloride (CMO) to generate a PbTx-2-CMO hapten and reacted with succinic anhydride (HS) to generate the PbTx-2-HS hapten. These haptens were conjugated to keyhole limpet hemocyanin (KLH) and bovine serum albumin (BSA) to prepare immunogen and coating antigen reagents, respectively, using the active ester method. After immunization and cell fusion, a broad-spectrum monoclonal antibody (mAb) termed mAb 1D3 was prepared. The 50% inhibitory concentration (IC50) values of the icELISA for PbTx-2, PbTx-1, and PbTx-3 were 60.71, 52.61, and 51.83 μg/kg, respectively. Based on the broad-spectrum mAb 1D3, an icELISA was developed to determine brevetoxin levels. Using this approach, the limit of detection (LOD) for brevetoxin was 124.22 μg/kg and recoveries ranged between 89.08% and 115.00%, with a coefficient of variation below 4.25% in oyster samples. These results suggest that our icELISA is a useful tool for the rapid monitoring of brevetoxins in oyster samples.
