Label-free differentiation and quantification of ricin, abrin from their agglutinin biotoxins by surface plasmon resonance
Label-free differentiation and quantification of ricin, abrin from their agglutinin biotoxins by surface plasmon resonance
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DOI：
10.1016/j.talanta.2021.122860
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发表时间：
2021-10-23
期刊：
影响因子：
6.1
通讯作者：
Xie, Jianwei
中科院分区：
文献类型：
--
作者：
Luo, Li;Yang, Jiewei;Xie, Jianwei
在这里，我们描述了一个亲和分子导向的表面等离子体共振（SPR）免疫传感器的蓖麻毒素和相思豆毒素的结构高度相似的凝集素生物毒素的无标记，区分和定量。通过引入蛋白G作为亲和捕获分子，我们实现了一个完整的策略，包括（i）筛选单克隆抗体以夹心形式配对，（ii）区分定量与凝集素，（iii）确定活性与非活性生物毒素，以及（iv）在单个芯片上捕获的生物毒素的结构鉴定。通过免疫磁珠的富集步骤，我们可以准确地测量蓖麻毒素或相思豆毒素的浓度降低到0.6 ng/mL（10 pM）在不同的复杂基质，如甜菊糖，蛋白粉，和人血浆，线性范围为两个或三个数量级，并满意的回收率。然后，我们对蓖麻子和野豌豆的混合粗提物以及禁化武组织第四次生物毒素演习的真实的样品进行了差异定量，以证明其实际可用性。首次提供了通过非共价分子表面直接从Protein G亲和芯片上获得的SPR质谱证据，为粗提物的结构鉴定提供了明确的依据。
Here we describe an affinity molecule-directed surface plasmon resonance (SPR) immunosensor for a label-free, differentiation and quantification of ricin and abrin from their structural highly like agglutinin biotoxins. By an introduction of protein G as the affinity capturing molecule, we fulfilled a complete strategy contains (i) screening monoclonal antibodies to be paired in a sandwiched format, (ii) differentiate quantification from the agglutinin, (iii) ascertain of active from inactive biotoxin, and (iv) structural identification of captured biotoxins on a single chip. By the aid of an enrichment step from immunomagnetic beads, we could accurately measure ricin or abrin with a concentration lowered to 0.6 ng/mL (10 pM) in different complex matrices such as stevia, protein powder, and human plasma, with linear ranges of two or three orders of magnitude, and satisfied recovery. We then differentially quantified the mixed crude extracts from castor beans and jequirity peas, and real samples from the fourth OPCW biotoxin exercise to prove the practical availability. We further provided a SPRmass spectrometric evidence directly obtained from Protein G affinity chip via a noncovalent molecule surface for the first time for definitely structural identification for crude extracts.