中文摘要：因为三聚氰胺有害，严禁在食品中掺入，但仍有人将三聚氰胺添加到乳制品中，伪造营养成分。本研究引入噬菌体展示技术来发现识别三聚氰胺的生物受体。采用两种半抗原和两种噬菌体展示方法，通过交叉检查彼此获得的结果来提高该技术的特异性。将6-氨基己酸与2-氯-4,6-二氨基-1,3,5-三嗪分别作为间隔区和表位位点反应合成三聚氰胺半抗原，并将其作为载体装饰在牛血清白蛋白(BSA)上，制备BSA半抗原。基于这些半抗原，对噬菌体展示肽进行测序鉴定，根据频率和相似度筛选出5个肽生物受体。采用氧化石墨烯量子点(GOQD)-Hg2+淬灭体系对所筛选肽进行简单验证，并测定其选择性和灵敏度。简而言之，三聚氰胺-hg2+配合物很容易通过7C-7C叠加在GOQD表面，从而通过电荷转移猝灭GOQD的荧光。但是，如果所筛选肽与三聚氰胺具有亲和力，在加入生物受体后，由于阻碍了三聚氰胺- hg2+复合物的形成，减弱的荧光强度会恢复。通过这一方法，本研究成功找出了有效的肽序列，并用于电化学阻抗谱系统，对三聚氰胺具有较高的灵敏度，而检出限较低。
外文摘要：Blending melamine into foodstuffs is prohibited, but despite its harmfulness, melamine is still added to dairy products for falsifying nutritional composition. In this study, to discover the bioreceptor recognizing melamine, phage display was introduced. For the technique, two types of haptens and two kinds of phage display methods were used to improve its specificity through cross-checking each acquired result. The melamine hapten was synthesized by reacting 6-aminohexanoic acid with 2-chloro-4,6-diamino-1,3,5-triazine, which function as a spacer and epitope site, respectively, and hapten was decorated on the bovine serum albumin (BSA) as a carrier material to manufacture BSA-hapten. Based on these haptens, the phage-displayed peptides are identified by sequencing, and five peptide bioreceptors were selected according to the frequency and similarity. To construct the simple verification procedure for the discovered peptides, graphene oxide quantum dot (GOQD)-Hg2+ quenching system was adopted, and their selectivity and sensitivity were determined. Briefly, the melamine-Hg2+ complex could easily be attached on the surface of GOQD through 7C-7C stacking, thus, the fluorescence of GOQD is quenched via charge transfer. However, if the screened peptides possess an affinity to melamine, the diminished fluorescence intensity will be recovered after adding the bioreceptor to the system resulting from hindering the formation of the melamine-Hg2+ complex. We successfully figured out the valid peptide sequences among the candidates through this approach, and the best hit was applied to the electrochemical impedance spectroscopic system. As a result, it shows a high sensitivity to the melamine with a low detection limit.
外文关键词：Phage display；Melamine measurement；Graphene oxide quantum dot； Bioreceptor；Fluorescence sensor；Electrochemical sensor
作者：Park, CY；Kim, MW；Park, KY；Baek, SH；Stephanie, R；Ghosh, S；Chun, HS；Huh, YS；Kim, JK；Park, JP；Park, TJ
作者单位：Chung Ang Univ；Inha Univ；Hong Kong Univ Sci & Technol
期刊名称：SENSORS AND ACTUATORS B-CHEMICAL
期刊影响因子：7.1
出版年份：2021
出版刊次：330
点击下载：通过噬菌体展示筛选三聚氰胺特异性生物受体，构建其基于纳米复合物猝灭的验证方法，并将筛选到的生物受体应用于三聚氰胺的电化学检测