Precipitate-enhanced SAW immunosensor for sensitive monitoring of mite allergens
Precipitate-enhanced SAW immunosensor for sensitive monitoring of mite allergens
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DOI：
10.1016/j.snb.2019.05.056
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发表时间：
2019-10
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影响因子：
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通讯作者：
Koji Toma;Koki Oishi;Misato Kato;Kana Kurata;Naoyuki Yoshimura;T. Arakawa;H. Yatsuda;K. Kanamori;K. Mitsubayashi
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文献类型：
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作者：
Koji Toma;Koki Oishi;Misato Kato;Kana Kurata;Naoyuki Yoshimura;T. Arakawa;H. Yatsuda;K. Kanamori;K. Mitsubayashi
利用辣根过氧化物酶（HRP）催化反应产生的酶解产物提高声表面波（SAW）免疫传感器监测空气中屋尘螨（HDM）过敏原的灵敏度。比较了4-氯-1-萘酚（4CN）、3，3 ′-二氨基联苯胺（DAB）和3，3 ′，5，5 ′-四甲基联苯胺（TMB）3种候选底物的信号放大和快速重复测定的适用性。将每种底物溶液施加到传感器表面，其中粉尘螨（Dermatophagoides farinae）的第2组HDM变应原（Der f2）被捕获抗体和HRP缀合的检测抗体夹在中间。从HRP催化反应的沉淀产物诱导的传感器输出和通过不同pH试剂再生后的传感器信号恢复，分别评估信号放大和传感器再生速率。结果表明，4CN在使用调节至pH 13的NaOH的三种基底中显示出最高的表面再生速率以及传感器输出的第二大增强。4CN的使用使得能够相对于没有HRP催化反应的传感器输出，对于1 ng/mLDer f2，将传感器输出提高13倍。沉淀增强的SAW免疫传感器的检测限为35 pg/mL（相当于2.3 pM），这是1.4倍和206倍优于ELISA和SAW免疫传感器没有沉淀，分别。这表明，在SAW免疫传感器的最佳条件下开发4CN实现了HDM过敏原的超灵敏监测。
Precipitates produced via horseradish peroxidase (HRP-) catalyzed reaction was exploited to enhance the sensitivity of a surface acoustic wave (SAW) immunosensor for monitoring airborne house dust mite (HDM) allergens. Three candidate substrates, 4-chloro-1-naphthol (4CN), 3,3′-diaminobenzidine (DAB) and 3,3′,5,5′-tetramethylbenzidine (TMB), were systematically compared in terms of signal amplification and suitability to rapid and repeated measurement. Each substrate solution was applied to the sensor surface where group 2 HDM allergens ofDermatophagoides farinae(Der f2) were sandwiched by capture and HRP-conjugated detection antibodies. The signal amplification and sensor regeneration rate were evaluated from the sensor output induced by precipitated product of HRP-catalyzed reaction and the sensor signal recovery after the regeneration via various pH reagents, respectively. As a result, it was revealed that 4CN showed the highest surface regeneration rate among the three substrates with use of NaOH adjusted to pH 13 as well as the second largest enhancement of the sensor output. Use of 4CN enabled to enhance the sensor output by a factor of 13 for 1 ng/mLDer f2 with respect to that without the HRP-catalyzed reaction. The limit of detection of the precipitate-enhanced SAW immunosensor was 35 pg/mL (equivalent to 2.3 pM), which was 1.4-fold and 206-fold better than those of ELISA and the SAW immunosensor without a precipitate, respectively. These indicate that exploiting 4CN under the optimal conditions in the SAW immunosensor realizes ultra-sensitive monitoring of HDM allergens.