Department of Physical Chemistry and Institute of Electrochemistry, Universidad de Alicante, 03080 Alicante, Spain.
Faculty of Science and Engineering, School of Science and the Environment, Division of Chemistry and Environmental Science, Manchester Metropolitan University, Chester Street, Manchester M1 5GD, UK.
Biosens Bioelectron. 2016 Mar 15;77:1168-74. doi: 10.1016/j.bios.2015.11.005. Epub 2015 Nov 4.
l-lactate is an essential metabolite present in embryonic cell culture. Changes of this important metabolite during the growth of human embryo reflect the quality and viability of the embryo. In this study, we report a sensitive, stable, and easily manufactured electrochemical biosensor for the detection of lactate within embryonic cell cultures media. Screen-printed disposable electrodes are used as electrochemical sensing platforms for the miniaturization of the lactate biosensor. Chitosan/multi walled carbon nanotubes composite have been employed for the enzymatic immobilization of the lactate oxidase enzyme. This novel electrochemical lactate biosensor analytical efficacy is explored towards the sensing of lactate in model (buffer) solutions and is found to exhibit a linear response towards lactate over the concentration range of 30.4 and 243.9 µM in phosphate buffer solution, with a corresponding limit of detection (based on 3-sigma) of 22.6 µM and exhibits a sensitivity of 3417 ± 131 µAM(-1) according to the reproducibility study. These novel electrochemical lactate biosensors exhibit a high reproducibility, with a relative standard deviation of less than 3.8% and an enzymatic response over 82% after 5 months stored at 4 °C. Furthermore, high performance liquid chromatography technique has been utilized to independently validate the electrochemical lactate biosensor for the determination of lactate in a commercial embryonic cell culture medium providing excellent agreement between the two analytical protocols.
l-乳酸是胚胎细胞培养中一种必需的代谢物。这种重要代谢物在人类胚胎生长过程中的变化反映了胚胎的质量和活力。在这项研究中，我们报告了一种灵敏、稳定且易于制造的电化学生物传感器，用于检测胚胎细胞培养物介质中的乳酸。丝网印刷的一次性电极被用作电化学传感平台，用于实现乳酸生物传感器的小型化。壳聚糖/多壁碳纳米管复合材料被用于乳酸氧化酶的酶固定化。这种新型电化学乳酸生物传感器的分析效果是针对模型（缓冲）溶液中的乳酸传感进行探索的，发现在磷酸盐缓冲溶液中，对乳酸的线性响应范围为 30.4 和 243.9 µM，相应的检测限（基于 3-sigma）为 22.6 µM，根据重复性研究，灵敏度为 3417 ± 131 µAM(-1)。这些新型电化学乳酸生物传感器表现出很高的重现性，在 4°C 下储存 5 个月后，相对标准偏差小于 3.8%，酶反应超过 82%。此外，高效液相色谱技术已被用于独立验证电化学乳酸生物传感器在商业胚胎细胞培养基中对乳酸的测定，两种分析方案之间具有极好的一致性。
