Preparation and preliminary application of monoclonal antibody against α-zearalanol
WEI Wan-gui~1, WANG Li~1, BAI Yun-feng, GONG Yi~1, GU Ming~2, ZHU Jian~2, HAN Wei~2, GUAN Rong~2 ~1Shanghai CASB Biotechnology CO., LTD, Chinese Academy of Sciences, Shanghai 200233, ~2Shanghai Entry-Exit Inspection and Quarantine Bureau of the Peoples Republic of China, Shanghai 200135, China
AIM: To prepare monoclonal antibody (mAb) against α-zearalanol (α-ZER) and develop an immunoassay for the detection of α-ZER and its analogues residues in food derived from animal tissues. METHODS: α-ZER was conjugated to BSA as immunogen to immunize BALB/c mice and mAb were prepared by hybridoma technique. mAb’s characteristics (titer, Ig subclass, specificity and relative affinity) were identified by ELISA. Standard inhibitive cure was made and sensitivity of the mAb was identified. 37 samples derived from animal liver were detected for α-ZER residues by competitive ELISA established in the study. RESULTS: 8 hybridoma cell lines stably secreting anti-α-ZER mAb were obtained. The titer of one of them(4E5)was 5.142×10~7.The Ig subclass was IgG1.The mAb was specific for α-ZER and its analogues and had no cross-reactivity with other compounds.8 positive results were found from the 37 samples derived from animal liver which were negative detected by HPLC. CONCLUSION: Anti-α-ZER mAb has been prepared successfully. A rapid method using the mAb for detecting α-ZER and its analogues residues in animal tissues has been established.