Development of a novel antibody probe useful for domoic acid detection

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), an...

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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:	eng
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
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Summary:	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).
ISSN:	0956-5663 1873-4235