Sankiewicz Anna, Hermanowicz Adam, Grycz Artur, Łukaszewski Zenon, Gorodkiewicz Ewa
Faculty of Chemistry, Bioanalysis Laboratory, University of Bialystok, Bialystok, Poland.
Anal Methods. 2021 Feb 7;13(5):642-646. doi: 10.1039/d0ay02047h. Epub 2021 Jan 13.
Leptin is a hormone that has a fundamental role in the regulation of feeding and energy balance. A developed specific SPRi immunosensor for leptin may be a new tool for leptin determination in blood plasma. The immunosensor consists of rabbit anti-leptin antibody immobilized on a gold chip via cysteamine linker, using the EDC/NHS protocol. Non-fluidic array SPRi is used for analytical signal formation. Under optimized conditions, the linear response range of the immunosensor covers concentrations from 0.23 to 5 ng mL. The LOD of the immunosensor is 0.07 ng mL, and the LOQ is 0.23 ng mL. The precision of measurement depends on leptin concentration, and is between 9.1% and 2.2%. Recoveries of the leptin spike are between 97% and 110%. The immunosensor and related analytical method were validated by parallel determination of leptin in series of plasma from children suffering from malnutrition and a control group, using Enzyme-Linked Immunosorbent Assay (ELISA) and SPRi. Pearson's correlation coefficient was equal to 0.991. The developed immunosensor and related method are more direct, faster and much simpler than ELISA.
瘦素是一种在调节进食和能量平衡中起重要作用的激素。一种开发出的用于瘦素的特异性表面等离子体共振免疫传感器（SPRi immunosensor）可能是测定血浆中瘦素的一种新工具。该免疫传感器由通过半胱胺连接子固定在金芯片上的兔抗瘦素抗体组成，采用1-乙基-3-(3-二甲氨基丙基)碳二亚胺盐酸盐/ N-羟基琥珀酰亚胺（EDC/NHS）方案。非流体阵列表面等离子体共振免疫传感器用于分析信号的形成。在优化条件下，免疫传感器的线性响应范围覆盖0.23至5 ng/mL的浓度。免疫传感器的检测限为0.07 ng/mL，定量限为0.23 ng/mL。测量精度取决于瘦素浓度，在9.1%至2.2%之间。瘦素加标回收率在97%至110%之间。通过使用酶联免疫吸附测定法（ELISA）和表面等离子体共振免疫传感器（SPRi）对营养不良儿童系列血浆和对照组血浆中的瘦素进行平行测定，验证了该免疫传感器及相关分析方法。皮尔逊相关系数等于0.991。所开发的免疫传感器及相关方法比ELISA更直接、更快且更简单。
