Silicon nitride sugar chips for detection of Ricinus communis proteins and Escherichia coli O157 Shiga toxins
Silicon nitride sugar chips for detection of Ricinus communis proteins and Escherichia coli O157 Shiga toxins
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用于检测蓖麻蛋白和大肠杆菌 O157 志贺毒素的氮化硅糖芯片
DOI：
10.1016/j.ab.2019.06.002
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发表时间：
2019
影响因子：
2.9
通讯作者：
Yoshihiro Nishida
中科院分区：
文献类型：
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作者：
Daiki Tanaka;Hirotaka Uzawa;Takehiro Nagatsuka;Yuki Oba;Atsunori Hiratsuka;Ken-ichi Tayama;Toshio Yoshida;Yasuo Seto;Hirofumi Dohi;Yoshihiro Nishida
设计并合成了具有氨基-三甘醇或叠氮基-三甘醇的乳糖苷，并将这两种衍生物固定在氮化硅（SiN）表面上。当点击反应应用于固定的叠氮基糖，蓖麻凝集素（RCA 120）被检测到具有较高的响应，通过反射干涉光谱（RIfS）。当N-羟基琥珀酰亚胺（NHS）方法应用于糖固定化，响应小于点击一。作为阴性对照的牛血清白蛋白（BSA）的反应可以忽略不计，但通过点击法制备的乳糖-SiN芯片比NHS方法制备的芯片更有效地抑制了非特异性结合。接下来，我们检查了通过点击反应制备的抗体固定化SiN芯片。然而，检测响应低于乳糖-SiN芯片的检测响应，这意味着通过点击反应的糖-芯片优于抗体-芯片上级。最后，为了检测大肠杆菌O 157：H7中的滋贺毒素，合成了含叠氮基三甘醇的球三糖（Gb 3），并通过点击反应将其固定在SiN芯片上。Gb 3-SiN芯片使我们能够检测浓度低于100 ng/mL的毒素。RCA 120、马革兰氏阳性菌、金雀花凝集素和BSA对Gb 3-SiN芯片无反应，显示了对该毒素的高度特异性。
Lactosides having either an amino-triethylene glycol or an azido-triethylene glycol were designed and synthesized, and the two derivatives were immobilized onto silicon nitride (SiN) surfaces. When a click reaction was applied for the immobilization of the azido-sugar, aRicinus communislectin (RCA120) was detected with a higher response by reflectometric interference spectroscopy (RIfS). When anN-hydroxysuccinimide (NHS) method was applied for the sugar immobilization, the response was less than that of the click one. The response of bovine serum albumin (BSA) as the negative control was negligible, but the lactose-SiN chip prepared by the click method suppressed nonspecific binding more effectively than did the chip from the NHS method. Next, we examined an antibody-immobilized SiN chip prepared by the click reaction. The detection response was, however, lower than that of the lactose-SiN chip, meaning that the sugar-chip by the click reaction was superior to the antibody-chip. Finally, to detect Shiga toxins fromEscherichia coliO157:H7, globotrisaccharide (Gb3) with an azido-triethylene glycol was synthesized and immobilized onto the SiN chip by the click reaction. The Gb3-SiN chips enabled us to detect the toxins at concentrations less than 100 ng/mL. RCA120, horse gram, gorse lectins and BSA showed no response to the Gb3-SiN chip, showing a high specificity for the toxin.