Selected landscape phage probe as selective recognition interface for sensitive total prostate-specific antigen immunosensor
Selected landscape phage probe as selective recognition interface for sensitive total prostate-specific antigen immunosensor
复制标题
选择景观噬菌体探针作为敏感总前列腺特异性抗原免疫传感器的选择性识别界面
DOI：
10.1016/j.bios.2018.01.046
复制
发表时间：
2018-05-30
影响因子：
12.6
通讯作者：
Liu, Aihua
中科院分区：
文献类型：
--
作者：
Han, Lei;Xia, Hongqi;Liu, Aihua
总前列腺特异性抗原（t-PSA）水平通常被认为是前列腺癌的关键指标。这里，针对 t-PSA 的噬菌体探针选自 f8/8 景观噬菌体库。经过三轮生物淘选，分离出四个 t-PSA 结合噬菌体克隆，并通过 DNA 测序进行鉴定。根据噬菌体捕获测定，展示融合肽 ATRSANGM 的噬菌体克隆对 t-PSA 显示出最高的亲和力和特异性。随后，t-PSA 特异性噬菌体在夹心酶联免疫吸附测定 (ELISA) 和微分脉冲伏安法 (DPV) 测定系统中用作 t-PSA 捕获探针。两种测定方法对于真实血清样品分析都显示出高特异性和可接受的可靠性。相比之下，DPV法比ELISA法（3.3-330 ng mL(-1)和1.6 ng mL(-1)）具有更宽的线性范围（0.01-100 ng mL(-1)）和更低的检出限（3 pg mL(-1)）。此外，DPV系统在实际样品检测中与权威方法的差异较小。令人兴奋的是，基于噬菌体探针的DPV免疫传感器对t-PSA的检测表现出高灵敏度，LOD达到pg水平，远低于报道的基于抗体的免疫传感器的值（通常高于0.1 ng mL(-1)）。由于生物相容性、多价性、稳定性和高结构同质性，t-PSA特异性景观噬菌体被证明是生物传感器中的一种新型特异性界面。
The level of total prostate-specific antigen (t-PSA) is generally known as the key index of prostate cancer. Here, phage probes against t-PSA were selected from f8/8 landscape phage library. After three rounds of biopanning, four t-PSA-binding phage clones were isolated and identified by the DNA sequencing. Based on the phage capture assay, the phage clone displaying the fusion peptide ATRSANGM showed highest affinity and specificity against t-PSA. Subsequently, the t-PSA-specific phage was used as t-PSA capture probe in a sandwich enzyme linked immunosorbent assay (ELISA) and differential pulse voltammetry (DPV) assay systems. Both assay methods showed high specificity and acceptable reliability for real serum samples analysis. By comparison, DPV method showed wider linear range (0.01-100 ng mL(-1)) and lower limit of detection (3 pg mL(-1)) than those (3.3-330 ng mL(-1) and 1.6 ng mL(-1)) of ELISA. Moreover, DPV system showed smaller distinction to the authoritative method in real samples assay. Excitingly, the phage probe based DPV immunosensor showed high sensitivity for the detection of t-PSA and LOD achieved the pg level, which was far lower than those values (usually above 0.1 ng mL(-1)) for reported immunosensors based on antibodies. Due to the biocompatibility, multivalency, stability, and high structural homogeneity, the t-PSA-specific landscape phage demonstrates as a novel specific interface in biosensors.