Ricin detection using phage displayed single domain antibodies.
Ricin detection using phage displayed single domain antibodies.
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DOI：
10.3390/s90100542
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发表时间：
2009
期刊：
影响因子：
--
通讯作者：
Anderson GP
中科院分区：
文献类型：
--
作者：
Goldman ER;Liu JL;Bernstein RD;Swain MD;Mitchell SQ;Anderson GP
将噬菌体展示的单域抗体 (sdAb) 与单体可溶性表达的 sdAb 和美洲驼多克隆抗体进行比较，以检测蓖麻毒素。 SdAb 由源自骆驼科动物纯重链抗体 (HcAb) 的可变结构域组成。尽管 HcAb 缺乏可变轻链，但它们及其衍生物 sdAb 能够以高亲和力结合抗原。 sdAb 的小尺寸（~16 kDa）虽然在许多方面都有优势，但限制了可以掺入的标签数量。合并多个标签的能力是报告元素的一个有益属性。恰巧的是，sdAb 通常是使用噬菌体展示方法来选择的。使用噬菌体 M13 上展示的 sdAb 作为报告元件，可以整合大量标签。我们已经通过酶联免疫吸附测定 (ELISA) 和 Luminex 流体阵列测定证明了 sdAb 和噬菌体展示的 sdAb 用于检测蓖麻毒素的用途。噬菌体展示的 sdAb 在 ELISA 和 Luminex 测定中对蓖麻毒素的检测效果提高了五到十倍，检测限分别为 1 ng/mL 和 64 pg/mL。噬菌体展示的 sdAb 在硝化纤维点印迹测定（一种常用于表位作图的方法）中可视化与靶标的结合也显着更有效。
Phage-displayed single domain antibodies (sdAb) were compared to monomeric solubly expressed sdAb and llama polyclonal antibodies for the detection of ricin. SdAb are comprised of the variable domain derived from camelid heavy chain only antibodies (HcAb). Although HcAb lack variable light chains, they as well as their derivative sdAb are able to bind antigens with high affinity. The small size of sdAb (∼16 kDa), while advantageous in many respects, limits the number of labels that can be incorporated. The ability to incorporate multiple labels is a beneficial attribute for reporter elements. Opportunely, sdAb are often selected using phage display methodology. Using sdAb displayed on bacteriophage M13 as the reporter element gives the potential for incorporating a very high number of labels. We have demonstrated the use of both sdAb and phage- displayed sdAb for the detection of ricin using both enzyme linked immunosorbent assays (ELISAs) and Luminex fluid array assays. The phage-displayed sdAb led to five to ten fold better detection of ricin in both the ELISA and Luminex assays, resulting in limits of detection of 1 ng/mL and 64 pg/mL respectively. The phage-displayed sdAb were also dramatically more effective for the visualization of binding to target in nitrocellulose dot blot assays, a method frequently used for epitope mapping.
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影响因子：
2.2
作者：
Maass, David R.;Sepulveda, Jorge;Shoemaker, Charles B.
通讯作者：
Shoemaker, Charles B.
影响因子：
8
作者：
Dumoulin, M;Conrath, K;Matagne, A
通讯作者：
Matagne, A
影响因子：
4.8
作者：
Newton, Jessica R.;Kelly, Kimberly A.;Deutscher, Susan L.
通讯作者：
Deutscher, Susan L.
影响因子：
--
作者：
Fulton, R. E.;Thompson, H. G.
通讯作者：
Thompson, H. G.
影响因子：
7.4
作者：
Goldman, Ellen R.;Anderson, George P.;Conway, Jerry;Sherwood, Laura J.;Fech, Melissa;Vo, BaoHan;Liu, Jinny L.;Hayhurst, Andrew
通讯作者：
Hayhurst, Andrew