School of Chemistry and Physics, Faculty of Science, Queensland University of Technology, 2 George Street, Brisbane, 4001, Australia.
School of Chemistry and Physics, Faculty of Science, Queensland University of Technology, 2 George Street, Brisbane, 4001, Australia.
Talanta. 2022 Jan 1;236:122879. doi: 10.1016/j.talanta.2021.122879. Epub 2021 Sep 11.
We present a sensitive label-free surface enhanced Raman spectroscopy (SERS) method for the discrimination between the recombinant and endogenous human Erythropoietin (EPO) isoforms. The proposed methodology comprises a lectin-functionalised extractor chip for the extraction of the recombinant human EPO (rhuEPO) and the endogenous EPO (enEPO) from blood plasma. The disulfide bond molecular structure of the purified isoforms was modified to chemisorb the biomolecules onto a SERS substrate in a unified orientation, thus maximizing the reproducibility and sensitivity of the SERS measurements. The acquired SERS spectra of the EPO isoforms showed diagnostic Raman bands that allowed for the discrimination between rhuEPO and enEPO. The method was also used for the SERS quantification of rhuEPO and enEPO down to 0.1 pM and 0.5 pM, respectively. The SERS determination of the protein isoforms was cross validated against ELISA. The new SERS method has strong potential for the rapid screening of rhuEPO doping in athletes and for the therapeutic drug monitoring of rhuEPO treatment in cancer patients.
我们提出了一种灵敏的无标记表面增强拉曼光谱（SERS）方法，用于区分重组和内源性人促红细胞生成素（EPO）同工型。该方法包括一个凝集素功能化的提取器芯片，用于从血浆中提取重组人 EPO（rhuEPO）和内源性 EPO（enEPO）。纯化同工型的二硫键分子结构被修饰，以使生物分子在 SERS 基底上以统一的取向化学吸附，从而最大限度地提高 SERS 测量的重现性和灵敏度。EPO 同工型的获得的 SERS 光谱显示出诊断性的 Raman 带，允许区分 rhuEPO 和 enEPO。该方法还用于 SERS 定量 rhuEPO 和 enEPO，分别低至 0.1 pM 和 0.5 pM。SERS 对蛋白质同工型的测定与 ELISA 交叉验证。新的 SERS 方法具有快速筛查运动员中 rhuEPO 兴奋剂和癌症患者中 rhuEPO 治疗的治疗药物监测的强大潜力。
