基于氧化石墨烯的表面等离子体共振生物传感器同时检测妊娠相关血浆蛋白-A 和 -A2。
Simultaneous Real-Time Detection of Pregnancy-Associated Plasma Protein-A and -A2 Using a Graphene Oxide-Based Surface Plasmon Resonance Biosensor.
机构信息
Department of Obstetrics and Gynecology, Mackay Memorial Hospital, Taipei, Taiwan.
Institute of Electro-Optical Science and Technology, National Taiwan Normal University, Taipei, Taiwan.
出版信息
Int J Nanomedicine. 2020 Mar 26;15:2085-2094. doi: 10.2147/IJN.S237938. eCollection 2020.
BACKGROUND
Pregnancy-associated plasma protein-A and -A2 (PAPP-A and -A2) are principally expressed in placental trophoblasts and play a critical role in the regulation of fetal and placental growth. PAPP-A2 shares 45% amino acid similarity with PAPP-A. This study aimed to investigate the efficacy of real-time detection of PAPP-A and PAPP-A2 using a novel surface plasmon resonance (SPR) biosensor based on graphene oxide (GO).
METHODS
Traditional SPR and GO-based SPR chips were fabricated to measure PAPP-A and PAPP-A2 concentrations. We compared SPR response curves of PAPP-A and PAPP-A2 between traditional SPR and GO-SPR biosensors. We also performed interference tests and specificity analyses among PAPP-A, PAPP-A2, and mixed interference proteins.
RESULTS
The time to detect PAPP-A and PAPP-A2 was about 150 seconds with both traditional SPR and GO-SPR biosensors. Approximately double SPR angle shifts were noted with the GO-SPR biosensor compared to the traditional SPR biosensor at a PAPP-A and PAPP-A2 concentration of 5 μg/mL. The limit of detection of the GO-SPR biosensor was as low as 0.5 ng/mL for both PAPP-A and PAPP-A2. Interference testing revealed that almost all of the protein bonded on the GO-SPR biosensor with anti-PAPP-A from the mixture of proteins was PAPP-A, and that almost no other proteins were captured except for PAPP-A2. However, the SPR signal of PAPP-A2 (5.75 mdeg) was much smaller than that of PAPP-A (13.76 mdeg). Similar results were noted with anti-PAPP-A2, where almost all of the protein bonded on the GO-SPR biosensor was PAPP-A2. The SPR signal of PAPP-A (5.17 mdeg) was much smaller than that of PAPP-A2 (13.94 mdeg).
CONCLUSION
The GO-SPR biosensor could distinguish PAPP-A and PAPP-A2 from various mixed interference proteins with high sensitivity and specificity. It could potentially be used to measure PAPP-A and PAPP-A2 in clinical blood samples during pregnancy.
背景
妊娠相关血浆蛋白 A 和 -A2（PAPP-A 和 -A2）主要在胎盘滋养细胞中表达，在调节胎儿和胎盘生长中发挥关键作用。PAPP-A2 与 PAPP-A 具有 45%的氨基酸相似性。本研究旨在探讨基于新型氧化石墨烯（GO）的表面等离子体共振（SPR）生物传感器实时检测 PAPP-A 和 PAPP-A2 的效果。
方法
传统 SPR 和基于 GO 的 SPR 芯片用于测量 PAPP-A 和 PAPP-A2 的浓度。我们比较了传统 SPR 和 GO-SPR 生物传感器的 PAPP-A 和 PAPP-A2 的 SPR 响应曲线。我们还对 PAPP-A、PAPP-A2 和混合干扰蛋白之间的干扰试验和特异性分析进行了研究。
结果
传统 SPR 和 GO-SPR 生物传感器检测 PAPP-A 和 PAPP-A2 的时间约为 150 秒。在 PAPP-A 和 PAPP-A2 浓度为 5μg/mL 时，GO-SPR 生物传感器的 SPR 角度变化约为传统 SPR 生物传感器的两倍。GO-SPR 生物传感器对 PAPP-A 和 PAPP-A2 的检测限低至 0.5ng/mL。干扰试验表明，与混合物中的其他蛋白相比，与抗 PAPP-A 结合的几乎所有蛋白都是 PAPP-A，除了 PAPP-A2 外，没有其他蛋白被捕获。然而，PAPP-A2 的 SPR 信号（5.75mdeg）明显小于 PAPP-A（13.76mdeg）。用抗 PAPP-A2 进行的类似实验中，与 GO-SPR 生物传感器结合的几乎所有蛋白都是 PAPP-A2。PAPP-A 的 SPR 信号（5.17mdeg）明显小于 PAPP-A2（13.94mdeg）。
结论
GO-SPR 生物传感器可以高灵敏度和特异性地区分 PAPP-A 和 PAPP-A2 与各种混合干扰蛋白。它有望用于测量妊娠期间临床血样中的 PAPP-A 和 PAPP-A2。
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