*Article* **Preparation of Monoclonal Antibody against Pyrene and Benzo [a]pyrene and Development of Enzyme-Linked Immunosorbent Assay for Fish, Shrimp and Crab Samples**

**Shuangmin Wu †, Huaming Li †, Xiaoyang Yin, Yu Si, Liangni Qin, Hongfei Yang, Jiaxu Xiao and Dapeng Peng \***

National Reference Laboratory of Veterinary Drug Residues (HZAU) and MOA Key Laboratory for the Detection of Veterinary Drug Residues in Foods, Huazhong Agricultural University, Wuhan 430070, China **\*** Correspondence: pengdapeng@mail.hzau.edu.cn; Tel.: +86-27-8728-7165; Fax: +86-27-8767-2232

† These authors contributed equally to this work.

**Abstract:** Polycyclic aromatic hydrocarbons (PAHs) are significant environmental and food pollutants that can cause cancer. In this work, a specific monoclonal antibody (mAb) to identify pyrene (PYR) and benzo [a]pyrene (BaP) was prepared, and an indirect competitive enzyme-linked immunoassay (ic-ELISA) was established to detect PYR and BaP residues in living aquatic products for the first time. The effects of complete antigens with different coupling ratios on the production of high-sensitivity mAb was explored. Under the optimal conditions, the IC50 value was 3.73 ± 0.43 μg/L (n = 5). The limits of detection (LODs) for PYR and BaP in fish, shrimp, and crab ranged from 0.43 to 0.98 μg/L. The average recoveries of the spiked samples ranged from 81.5–101.9%, and the coefficient of variation (CV) was less than 11.7%. The validation of the HPLC-FLD method indicated that the ELISA method set up in this experiment provided a trustworthy tool for PAHs residues detection in aquatic products.

**Keywords:** pyrene; benzo [a]pyrene; monoclonal antibody; enzyme-linked immunoassay; aquatic product
