Centre for Biomarker Research and Development, Australian Institute for Bioengineering and Nanotechnology (AIBN), The University of Queensland, Brisbane QLD 4072, Australia.
Centre for Biomarker Research and Development, Australian Institute for Bioengineering and Nanotechnology (AIBN), The University of Queensland, Brisbane QLD 4072, Australia.
Biosens Bioelectron. 2014 May 15;55:417-22. doi: 10.1016/j.bios.2013.12.043. Epub 2013 Dec 27.
The time and costs associated with monoclonal antibody production limit the potential for portable diagnostic devices to penetrate the market. Replacing the antibody with a low-cost alternate affinity reagent would reduce the costs of diagnostic development and use, and lead to new portable diagnostic devices towards many diseases. Herein, we present low-cost affinity reagents, nano-yeast-scFv, on commercially available, inexpensive, and portable screen-printed electrodes for the label-free electrochemical detection of Entamoeba histolytica cyst antigens. The biosensor was able to detect antigen at concentrations down to 10 pg mL(-1) in buffer with an inter-assay reproducibility of (% RSD, n=3) 4.1%. The applicability of two differently engineered nano-yeast-scFv to each specifically detect their cognant E. histolytica cyst antigens was demonstrated in a biological matrix derived from human stool. Because of the simple, inexpensive, and sensitive nature of this methodology, it may offer a low-cost alternative to immunosensors based on antibody-target recognition.
单克隆抗体生产的时间和成本限制了便携式诊断设备进入市场的潜力。用低成本的替代亲和试剂替代抗体将降低诊断开发和使用的成本,并为许多疾病开发出新型的便携式诊断设备。在此,我们提出了低成本的亲和试剂,纳米酵母-scFv,可用于市售的、廉价的、便携式的丝网印刷电极,用于无标记电化学检测溶组织内阿米巴包囊抗原。该生物传感器能够在缓冲液中检测到浓度低至 10 pg mL(-1)的抗原,在 3 次测定内的重现性为(% RSD,n=3)4.1%。两种不同工程化的纳米酵母-scFv 能够分别特异性地检测其识别的溶组织内阿米巴包囊抗原,这在源自人粪便的生物基质中得到了证明。由于该方法简单、廉价且灵敏,它可能为基于抗体-靶标识别的免疫传感器提供一种低成本替代方案。