Development of a novel antibody probe useful for domoic acid detection
Development of a novel antibody probe useful for domoic acid detection
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开发可用于软骨藻酸检测的新型抗体探针。
DOI：
10.1016/j.bios.2009.03.023
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发表时间：
2009-06-15
影响因子：
12.6
通讯作者：
Zhang, Jun-Hui
中科院分区：
文献类型：
--
作者：
Zhou, Yu;Zhang, Yuan-Yuan;Zhang, Jun-Hui
海洋毒素单克隆抗体（mAb）的产生为利用免疫学方法检测该类化合物提供了一种有价值的探针。然而，传统的单克隆抗体生成方法通常需要相对大量的标准物质（超过400 μ g小鼠（-1））和相对较长的免疫周期（超过6周）。在这里，我们报告了一种简单，廉价和快速的方案，用于生成软骨藻酸（DA）特异性单克隆抗体探针。该方法取Balb/C后足垫注射小鼠淋巴细胞，与小鼠骨髓瘤细胞SP2/0融合产生杂交瘤。这种制备DA单抗的方法有两个主要优点：(a)不需要大规模昂贵的抗原（一只小鼠仅需70 μ g抗原）；(b)免疫方案可在16天内完成。研究了单抗的一些特性，并开发了以单抗为探针的直接竞争性ELISA检测DA。磷酸盐缓冲盐水（PBS）的检出限为0.41 ng /(-1)，蓝贻贝（Mytilus edulis）的检出限为0.53 ng /(-1)。贻贝和PBS缓冲液中DA的回收率分别为94.8% ~ 105.1%和96.2% ~ 103.7%。因此，新开发的使用单抗的直接竞争性ELISA似乎是监测贝类DA的可靠和有用的方法（228字）。(C) 2009 Elsevier B.V.版权所有
The generation of monoclonal antibody (mAb) against marine toxins call 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 mu g 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 mu g 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). (C) 2009 Elsevier B.V. All rights reserved.