Portable immuno nano biosensor for multiple disease detection using 3D printed SPR technologies

Aparanji Poosarla, Jagannadha Rao Mokka, Kolla Venkata Atchuta Rao, Mano Pardhavi Dasari
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Abstract

The role of biosensors as point-of-care devices is well explored in recent years and their usage in the current market has been highly increased after the outbreak of the COVID pandemic. The point-of-care devices generally use sensing principles of electrochemical, immune fluorescence, microbial and electromagnetic. The sensor we developed uses the phenomenon of Surface Plasmon resonance with optic fiber technology which is a novel design to implement in point-of-care devices. With the latest innovations and improvements in 3D printing technologies the development and prototyping of these POC devices have been accelerated to the greatest potential. Here the entire device was prototyped using Creality Upgraded Ender 3 2021 model 3D printer where the 3D files were sliced in Ultimaker Cura software and printed with PLA filament by fused filament deposition modeling (FFDM). The device has a USB light and camera source on either side of the sensing chamber and a cassette with optic fiber coated with respective antibodies as a sensing platform. The device is connected to respective USB ports and introduced with the sample over a cassette and placed in the reaction chamber. The reaction occurs spontaneously which is captured by the webcam and recorded by the software. We tested the bare sensor with increased concentrations of glycerol after coating it with a silver mirror by a standard reproducible method of Tollens silver mirror. The sensor showed the best sensitivity with increasing concentrations of glycerol. The sensor also immobilized with target antibodies of pLDH parasite lactate dehydrogenase by non-specific binders just by adding target bio-analyte and receptor over silver mirror functionalized surface and tested with positive control antigen of malaria parasite antigen. The sensor showed the highest sensitivity at 50X concentration of antigen and least at 10x concentration stating maximum and minimum values for a limit of detection. This device using this principle could has wide functionalities to detect multiple biological samples by changing the antibodies to our respective analyte over the reaction chip for multiple times with low cost.

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利用3D打印SPR技术检测多种疾病的便携式免疫纳米生物传感器
近年来,生物传感器作为护理点设备的作用得到了很好的探索,在新冠肺炎疫情爆发后,它们在当前市场上的使用量大幅增加。护理点设备通常使用电化学、免疫荧光、微生物和电磁的传感原理。我们开发的传感器将表面等离子体共振现象与光纤技术相结合,这是一种在护理点设备中实现的新颖设计。随着3D打印技术的最新创新和改进,这些POC设备的开发和原型设计已经加速到最大潜力。在这里,使用Creality Upgraded Ender 3 2021型号3D打印机对整个设备进行原型化,其中3D文件在Ultimaker Cura软件中切片,并通过熔融细丝沉积建模(FFDM)用PLA细丝打印。该设备在传感室的两侧都有一个USB光源和摄像头,还有一个带光纤的盒子,光纤上涂有各自的抗体作为传感平台。该装置连接到各自的USB端口,并与样品一起通过盒引入并放置在反应室中。这种反应是自发发生的,由网络摄像头捕捉并由软件记录下来。我们通过Tollens银镜的标准可重复方法,在用银镜涂覆裸传感器后,用增加浓度的甘油对其进行了测试。随着甘油浓度的增加,传感器显示出最佳的灵敏度。仅通过在银镜功能化表面上添加靶生物分析物和受体,传感器也通过非特异性粘合剂用pLDH寄生虫乳酸脱氢酶的靶抗体固定,并用疟原虫抗原的阳性对照抗原进行测试。该传感器在抗原浓度为50倍时显示出最高灵敏度,在检测极限的最大值和最小值为10倍时显示最低灵敏度。这种使用这一原理的设备可以具有广泛的功能,通过在反应芯片上多次低成本改变我们各自分析物的抗体来检测多个生物样本。
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