Development of a novel antibody probe useful for domoic acid detection

• Published in: Biosensors & bioelectronics 2009-06, Vol.24 (10), p.3159-3163
• Main Authors: Zhou, Yu, Zhang, Yuan-Yuan, Shen, Qing-Feng, Lu, Shi-Ying, Ren, Hong-Lin, Li, Yan-Song, Liu, Zeng-Shan, Pan, Feng-Guang, Meng, Xian-Mei, Zhang, Jun-Hui
• Format: Article
• Language: English
• Subjects: Animals; Antibodies, Monoclonal; Biological and medical sciences; Biotechnology; Cell Line, Tumor; Domoic acid; Electrophoresis, Agar Gel; Enzyme-Linked Immunosorbent Assay - methods; Female; Fundamental and applied biological sciences. Psychology; Hybridoma cell line; Hybridomas; Immunization - methods; Immunological method; Kainic Acid - analogs & derivatives; Kainic Acid - analysis; Kainic Acid - immunology; Lymph node cell; Male; Marine toxin; Marine Toxins - analysis; Marine Toxins - immunology; Mice; Mice, Inbred BALB C; Monoclonal antibody; Mytilus edulis; Mytilus edulis - chemistry
• ISSN: 0956-5663; 1873-4235
• DOI: 10.1016/j.bios.2009.03.023

The generation of monoclonal antibody (mAb) against marine toxins can serve as a valuable probe to detect this kind of compounds by immunological methods. However, traditional approaches to mAb generation usually need a comparative large quantity of standard substance (more than 400 μg mouse −1), and a comparative long immunization period (more than 6 weeks). Here we report a simple, inexpensive and fast protocol for the generation of monoclonal antibody probe specific for domoic acid (DA). In the method, lymph node cells were harvested from the Balb/C mice of hind footpad injection and fused with murine myeloma cells SP2/0 for hybridoma generation. This method for the preparation of mAb for DA has two main advantages: (a) there is no need for large-scale expensive antigen (only 70 μg antigen for one mouse); (b) immunization protocol can be accomplished within 16 days. Some characteristics of the mAb were studied and a direct competitive ELISA for the detection of DA using the mAb as a probe was developed. The detection limit was 0.41 ng well −1 in phosphate buffered saline (PBS) and 0.53 ng well −1 in blue mussel Mytilus edulis. The recoveries of DA from mussel and PBS buffer were from 94.8% to 105.1% and from 96.2% to 103.7%, respectively. Thus, the newly developed direct competitive ELISA using the mAb appears to be a reliable and useful method for monitoring of DA in shellfish (228 words).