Amarnath Chellachamy Anbalagan, Jyoti Korram, Mukesh Doble, Shilpa N Sawant
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To further improve the sensor performance, hyper-branched polyethylenimine grafted poly amino aniline (PEI-g-PAANI) was used as the sensing interface, which enabled immobilization of higher amount of capture antibody. Detection of CEA in human blood serum coupled with wide linear range (0.5-50 ng/ml), good specificity, stability, reproducibility and low detection limit (10 pg/ml) signified the excellence of CUCDs based CEA immunosensor. CUCDs exhibited excitation wavelength dependent fluorescence property and showed strong blue emission under UV irradiation. MTT assay indicated that the material is not toxic towards human dental pulp stem cells (hDPSCs) and MG63 osteosarcoma cells (cell viability > 90%). 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引用次数: 0
摘要
癌症的早期诊断需要对血液中的癌症生物标志物进行灵敏而准确的检测。碳点(CD)与抗体、多肽或适配体的生物功能化在癌症诊断和癌症靶向治疗中发挥了重要作用。在此,我们利用与 HRP 结合物 CEA 抗体(CUCDs@CEAAb2)生物功能化的碳点作为检测探针,设计了一种用于检测血清中癌症生物标志物癌胚抗原(CEA)的生物传感器。CDs 是通过水热法将富含氮的生物质废物牛尿升级合成的。与传统的电化学夹心免疫分析法相比,基于 CDs 的检测探针的灵敏度提高了 3.5 倍。为了进一步提高传感器的性能,使用了超支化聚乙烯亚胺接枝聚氨基苯胺(PEI-g-PAANI)作为传感界面,从而固定了更多的捕获抗体。人血清中 CEA 的检测具有线性范围宽(0.5-50 ng/ml)、特异性好、稳定性好、重现性好和检测限低(10 pg/ml)等特点,这表明基于 CUCDs 的 CEA 免疫传感器非常出色。CUCDs 具有与激发波长相关的荧光特性,在紫外线照射下会发出强烈的蓝色荧光。MTT 试验表明,该材料对人牙髓干细胞(hDPSCs)和 MG63 骨肉瘤细胞无毒性(细胞存活率大于 90%)。本研究展示了一种将动物废弃物转化为具有成本效益的碳基功能纳米材料,用于临床检测癌症生物标志物的方法。
Bio-functionalized carbon dots for signaling immuno-reaction of carcinoembryonic antigen in an electrochemical biosensor for cancer biomarker detection.
Early diagnosis of cancer demands sensitive and accurate detection of cancer biomarkers in blood. Carbon dots (CDs) bio-functionalization with antibodies, peptides or aptamers have played significant role in cancer diagnosis and targeted cancer therapy. Herein, a biosensor for detection of cancer biomarker carcinoembryonic antigen (CEA) in blood serum has been designed using CDs bio-functionalized with HRP-conjugated CEA antibody (CUCDs@CEAAb2) as detection probe. CDs were synthesized by upscaling of cow urine, a nitrogen rich biomass waste, by hydrothermal method. Detection probe based on CDs resulted in 3.5 times higher sensitivity as compared to conventional electrochemical sandwich immunoassay. To further improve the sensor performance, hyper-branched polyethylenimine grafted poly amino aniline (PEI-g-PAANI) was used as the sensing interface, which enabled immobilization of higher amount of capture antibody. Detection of CEA in human blood serum coupled with wide linear range (0.5-50 ng/ml), good specificity, stability, reproducibility and low detection limit (10 pg/ml) signified the excellence of CUCDs based CEA immunosensor. CUCDs exhibited excitation wavelength dependent fluorescence property and showed strong blue emission under UV irradiation. MTT assay indicated that the material is not toxic towards human dental pulp stem cells (hDPSCs) and MG63 osteosarcoma cells (cell viability > 90%). The present study demonstrates a methodology for valorization of animal waste to a cost-effective carbon based functional nanomaterial for clinical detection of cancer biomarkers.