使用量身定制的重组抗体片段和噬菌体展示方法对未酰胺化的前胃泌素衍生肽（G17-Gly）进行生物测定：生物医学分析。,Analytical Methods

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使用量身定制的重组抗体片段和噬菌体展示方法对未酰胺化的前胃泌素衍生肽（G17-Gly）进行生物测定：生物医学分析。
Analytical Methods ( IF 2.7 ) Pub Date : 2020-04-16 , DOI: 10.1039/d0ay00627k
Deniz Sadighbayan _{1,

2} , Mohammad Reza Tohidkia ₃ , Tayebeh Mehdipour ₃ , Mohammad Hasanzadeh _{4,

5} , Ahmad Yari Khosroushahi _{1,

6}

Affiliation

在这项研究中，基于信号放大和量身定制的重组抗体技术，设计了四种新颖，灵敏的电化学测定G17-Gly的免疫传感器。抗G17-Gly抗体片段（即将对G17-Gly的N和C末端特异的scFv和VL固定在聚合物纳米复合材料上，该复合材料包括聚十六烷基三甲基溴化铵（P（CTAB））作为导电基质，壳聚糖（CS）作为生物相容剂和金纳米粒子（AuNPs）作为信号放大元件。AuNPs提供的高表面积和scFv / VL的小尺寸为将大量抗G17-Gly固定在检测人血浆样品中G17-Gly的电极表面上奠定了基础。在最佳条件下，设计的免疫传感器具有出色的分析能力，可检测和测定人血浆样品中的G17-Gly，线性范围为0.5 mM至0.05 pM，LLOQ为0.05 pM。免疫传感器的灵敏度顺序为Ag / 2-巯基乙醇/噬菌体，显示scFv / P（CTAB–CS）–AuNP / GE，Ag / 2-巯基乙醇/噬菌体，显示VL / P（CTAB–CS）–AuNP / GE，Ag / BSA / scFv / P（CTAB–CS）–AuNP / GE和Ag / BSA / VL / P（CTAB–CS ）–AuNP / GE。前述特征表明，所提出的免疫装置可以在生物学和临床诊断中用作鉴定不同癌生物标志物的可靠工具。

"点击查看英文标题和摘要"

Bio-assay of the non-amidated progastrin-derived peptide (G17-Gly) using the tailor-made recombinant antibody fragment and phage display method: a biomedical analysis.

In this research, four novel and sensitive immunosensors for electrochemical determination of G17-Gly were designed based on signal amplification and tailor-made recombinant antibody technology. Anti-G17-Gly antibody fragments (i.e. scFv and VL specific to the N- and C-terminal of G17-Gly) were immobilized onto a polymeric nanocomposite comprising poly cetyl trimethyl ammonium bromide (P(CTAB)) as the conductive matrix, chitosan (CS) as a biocompatible agent and gold nanoparticles (AuNPs) as the signal amplification element. The high surface area provided by AuNPs and the small size of scFv/VL establish the basis for immobilizing a high amount of the anti-G17-Gly on the surface of the electrode for detecting G17-Gly in human plasma samples. Under optimal conditions, the designed immunosensors provide an excellent analytical capability for detecting and determining G17-Gly in human plasma samples with a linear range from 0.5 mM to 0.05 pM and a LLOQ of 0.05 pM. The sensitivity order of the immunosensors was Ag/2-mercaptoethanol/phage displaying scFv/P(CTAB–CS)–AuNP/GE, Ag/2-mercaptoethanol/phage displaying VL/P(CTAB–CS)–AuNP/GE, Ag/BSA/scFv/P(CTAB–CS)–AuNP/GE, and Ag/BSA/VL/P(CTAB–CS)–AuNP/GE. The aforementioned characteristics demonstrate that the proposed immune-devices can be used in biological and clinical diagnosis as reliable tools for identifying different oncobiomarkers.

更新日期：2020-04-16

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