Detection of Abrin Holotoxin Using Novel Monoclonal Antibodies.
Detection of Abrin Holotoxin Using Novel Monoclonal Antibodies.
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DOI：
10.3390/toxins9120386
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发表时间：
2017-11-28
期刊：
影响因子：
4.2
通讯作者：
Brandon DL
中科院分区：
文献类型：
--
作者：
He X;Patfield S;Cheng LW;Stanker LH;Rasooly R;McKeon TA;Zhang Y;Brandon DL
相思子是核糖体失活蛋白家族的成员，由相思子植物产生。由于对人类和动物健康都有可能造成严重威胁，相思豆毒素被美国卫生与公众服务部列为选择性制剂。然而，尚未报道特异性针对完整相思豆毒素全毒素的免疫测定。在这项研究中，七个新的单克隆抗体（单克隆抗体），指定为ARAM-I通过ARAM-7已经开发。同种型分析表明这些mAb具有IgG 1、IgG 2a或IgG 2b重链和κ轻链。蛋白质印迹分析鉴定了两种相思豆毒素A链特异性mAb，A β-1和A β-2，以及四种B链特异性mAb（A β-3、A β-5、A β-6和A β-7）。以B链特异性的亲和素A-3为捕获剂，A链特异性的亲和素A-2为检测剂，建立了一种检测相思豆毒素和凝集素混合物的夹心酶联免疫吸附试验（ELISA）。ELISA是高度敏感的，并检测1 ng/mL的相思豆毒素全毒素在磷酸盐缓冲盐水，脱脂牛奶，和全脂牛奶，显着低于浓度，会对消费者造成健康问题。该ELISA还以非常低的背景信号检测植物提取物中的天然相思豆毒素。新的相思豆毒素单克隆抗体和ELISA应该是有用的检测这种强大的毒素在牛奶供应链和其他复杂的矩阵。
Abrin, a member of the ribosome-inactivating protein family, is produced by the Abrus precatorius plant. Having the potential to pose a severe threat to both human and animal health, abrin is classified as a Select Agent by the U.S. Department of Health and Human Services. However, an immunoassay that is specific for intact abrin holotoxin has not yet been reported. In this study, seven new monoclonal antibodies (mAbs), designated as Abrin-1 through Abrin-7 have been developed. Isotyping analyses indicate these mAbs have IgG1, IgG2a, or IgG2b heavy-chains and kappa light-chains. Western blot analyses identified two abrin A-chain specific mAbs, Abrin-1 and Abrin-2, and four B-chain specific mAbs (Abrin-3, -5, -6, and -7). A sandwich enzyme-linked immunosorbent assay (ELISA), capable of detecting a mixture of abrin isoforms and agglutinins was developed using B-chain specific Abrin-3 for capture and A-chain specific Abrin-2 as detector. The ELISA is highly sensitive and detects 1 ng/mL of the abrin holotoxin in phosphate-buffered saline, nonfat milk, and whole milk, significantly below concentrations that would pose a health concern for consumers. This ELISA also detects native abrin in plant extracts with a very low background signal. The new abrin mAbs and ELISA should be useful for detecting this potent toxin in the milk supply chain and other complex matrices.
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DOI：
10.1111/j.1432-1033.1991.tb16072.x
发表时间：
1991-06-15
期刊：
EUROPEAN JOURNAL OF BIOCHEMISTRY
影响因子：
--
作者：
WOOD, KA;LORD, JM;PIATAK, M
通讯作者：
PIATAK, M
DOI：
10.1073/pnas.0408526102
发表时间：
2005-07-12
影响因子：
11.1
作者：
Wein, LM;Liu, YF
通讯作者：
Liu, YF
影响因子：
2.9
作者：
Jang, David H.;Hoffman, Robert S.;Nelson, Lewis S.
通讯作者：
Nelson, Lewis S.
DOI：
10.2165/00139709-200322030-00002
发表时间：
2003-01-01
期刊：
Toxicological Reviews
影响因子：
--
作者：
Dickers, Kirsten J.;Bradberry, Sally M.;Vale, J. Allister
通讯作者：
Vale, J. Allister
影响因子：
3.5
作者：
OLSNES, S;PIHL, A
通讯作者：
PIHL, A