Shah Maitri, Kolhe Pratik, Roberts Akanksha, Shrikrishna Narlawar Sagar, Gandhi Sonu
DBT -National Institute of Animal Biotechnology (NIAB), Hyderabad 500032, Telangana, India.
DBT -National Institute of Animal Biotechnology (NIAB), Hyderabad 500032, Telangana, India.
Colloids Surf B Biointerfaces. 2022 Nov;219:112812. doi: 10.1016/j.colsurfb.2022.112812. Epub 2022 Aug 28.
The impact of uncontrolled antibiotic use in animals has subsequently led to emergence of antibiotic-resistant bacteria among humans due to consumption of animal by-products. Hence, to investigate antibiotic contamination in animal origin food products, we have developed a reduced graphene oxide (rGO) based immunosensor using fabricated electrode conjugated with anti-Penicillin antibody (rGO/Pen-Ab) for sensitive detection of Penicillin G. To execute this, Penicillin was first conjugated with Bovine Serum Albumin (BSA) which was confirmed via chromatographic, spectroscopic and electrophoretic-based techniques against both the in-house developed Penicillin conjugate (Pen-BSA) as well as the commercial Penicillin conjugate (Com-Pen-BSA). Further, we fabricated electrode based on one step synthesized rGO and immobilized with antibodies generated against Pen-BSA (Pen-Ab), and Com-Pen-BSA (Com-Pen-Ab), separately for detection of Penicillin. Each synthesis and conjugation step was confirmed by different spectroscopic methods. For efficient working of the electrode, various parameters were optimized using Voltammetry. The limit of detection for Penicillin G against Pen-Ab and Com-Pen-Ab was determined as 0.724 pM and 0.668 pM respectively and both displayed negligible cross reactivity against other β-lactam antibiotics (Cefalexin and Ampicillin). Furthermore, antibiotics were also detected in spiked milk, egg and meat samples and the electrode was evaluated for repeatability and storage stability. In conclusion, in-house developed Pen-Ab showed better sensitivity as compared to Com-Pen-Ab. The fabricated rGO/Pen-Ab biosensor shows future potential for rapid detection of penicillin and other β-lactam antibiotics for safe consumption of animal by-products in humans.
动物中抗生素的无节制使用，随后导致人类因食用动物副产品而出现抗生素抗性细菌。因此，为了研究动物源性食品中的抗生素污染情况，我们开发了一种基于还原氧化石墨烯（rGO）的免疫传感器，该传感器使用与抗青霉素抗体（rGO/Pen-Ab）共轭的预制电极，用于灵敏检测青霉素G。为此，首先将青霉素与牛血清白蛋白（BSA）共轭，通过色谱、光谱和基于电泳的技术，针对内部开发的青霉素共轭物（Pen-BSA）以及市售青霉素共轭物（Com-Pen-BSA）进行了确认。此外，我们基于一步合成的rGO制备了电极，并分别固定针对Pen-BSA（Pen-Ab）和Com-Pen-BSA（Com-Pen-Ab）产生的抗体，用于检测青霉素。每个合成和共轭步骤都通过不同的光谱方法进行了确认。为了使电极有效工作，使用伏安法对各种参数进行了优化。针对Pen-Ab和Com-Pen-Ab的青霉素G检测限分别确定为0.724 pM和0.668 pM，并且两者对其他β-内酰胺抗生素（头孢氨苄和氨苄青霉素）的交叉反应性可忽略不计。此外，还在加标的牛奶、鸡蛋和肉类样品中检测到了抗生素，并对电极的重复性和储存稳定性进行了评估。总之，与Com-Pen-Ab相比，内部开发的Pen-Ab表现出更好的灵敏度。所制备的rGO/Pen-Ab生物传感器在快速检测青霉素和其他β-内酰胺抗生素以确保人类安全食用动物副产品方面显示出未来潜力。
