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使用 S-乙酰巯基琥珀酸酐(SAMSA)修饰抗体对 Glypican-1 进行检测的生物传感器的高级制造。

Advanced fabrication of biosensor on detection of Glypican-1 using S-Acetylmercaptosuccinic anhydride (SAMSA) modification of antibody.

机构信息

Department of Chemical and Biomolecular Engineering, Case Western Reserve University, 10900 Euclid Ave, Cleveland, OH, 44106, USA.

Electronics Design Center, Case Western Reserve University, 10900 Euclid Ave, Cleveland, OH, 44106, USA.

出版信息

Sci Rep. 2018 Sep 10;8(1):13541. doi: 10.1038/s41598-018-31994-2.

Abstract

Glypican-1 (GPC-1) has been recognized as biomarker of pancreatic cancer. Quantification of GPC-1 level is also pivotal to breast cancer and prostate cancer's patients. We hereby report the first biosensor for GPC-1 detection. Instead of using crosslinking technique and surface immobilization of antibody, we applied a novel method for biosensor fabrication, using S-Acetylmercaptosuccinic anhydride (SAMSA) to modify the Anti-GPC-1 producing a thiol-linked Anti-GPC-1. The thiol-linked Anti-GPC-1 was then directly formed a single-layer antibody layer on the gold biosensor, minimizing the biosensor preparation steps significantly. Time of Flight Secondary Ions Mass Spectroscopy (TOF-SIMS) characterization verified the thiol-linked antibody layer and demonstrated a unique perspective for surface protein characterization. Differential pulse voltammetry (DPV) was applied to quantify GPC-1 antigen in undiluted human serum with a concentration range of 5,000 pg/µL to 100 pg/µL. The performance of this newly designed biosensor was also compared with modified self-assembled monolayer system fabricated biosensor, demonstrating the high-sensitivity and high-reproducibility of the SAMSA modified antibody based biosensor. This simple fabrication method can also expand to detection of other biomolecules. The simplified operation process shows great potential in clinical application development.

摘要

聚糖蛋白-1(GPC-1)已被认为是胰腺癌的生物标志物。定量检测 GPC-1 水平对于乳腺癌和前列腺癌患者也至关重要。我们在此报告了第一个用于检测 GPC-1 的生物传感器。我们采用了一种新的方法来制备生物传感器,而不是使用交联技术和抗体表面固定化,而是使用 S-乙酰巯基琥珀酸酐(SAMSA)修饰抗 GPC-1 产生硫醇连接的抗 GPC-1。然后,硫醇连接的抗 GPC-1 直接在金生物传感器上形成单层抗体层,大大简化了生物传感器的制备步骤。飞行时间二次离子质谱(TOF-SIMS)表征验证了硫醇连接的抗体层,并为表面蛋白特性提供了独特的视角。差分脉冲伏安法(DPV)用于在未稀释的人血清中定量检测 5000 pg/µL 至 100 pg/µL 浓度范围内的 GPC-1 抗原。与使用修饰的自组装单分子层系统制备的生物传感器相比,新设计的生物传感器的性能也进行了比较,证明了基于 SAMSA 修饰抗体的生物传感器具有高灵敏度和高重现性。这种简单的制造方法也可以扩展到其他生物分子的检测。简化的操作过程在临床应用开发中具有很大的潜力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d217/6131508/acd5c6b315fb/41598_2018_31994_Fig2_HTML.jpg

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