Surface-enhanced Raman probe for rapid nanoextraction and detection of erythropoietin in urine

Abstract

We present a surface-enhanced Raman probe (SERS) platform for the determination of a prohibited substance, recombinant erythropoietin (rEPO), in urine matrix, using nanoparticles as substrate. Rod-shaped gold nanoparticles (GNR) were modified with a Raman label and an antibody as SERS probe. We developed two SERS-based immunoassays for detection and quantification of rEPO in urine. In the first assay, rEPO was determined by a sandwich assay with gold surfaces and GNR. In the second assay, rEPO was extracted by using core shell-structured magnetic iron oxide gold nanoparticles, and again sandwich assay was performed by using GNR. We also demonstrated the ability of the proposed method to discriminate rEPO and urinary erythropoietin (uEPO). A good linear correlation was obtained between logarithms of rEPO concentrations in urine and Raman intensities within the range of 10-10 pg mL rEPO concentrations. Detection limits which are smaller than 0.1 pg mL levels were achieved owing to the high extractive performance of the nanoextraction techniques. Graphical Abstract Schematic represantation of surface-enhanced Raman probe for rapid nanoextraction and detection of erythropoietin.

Fe3O4@SiO2 Nanoparticles Functionalized with Gold and Poly(vinylpyrrolidone) for Bio-Separation and Sensing Applications

“…The ability of the Fe 3 O 4 @SiO 2 + Au + PVP nanoparticles to resolve peaks at low concentrations and demonstrate a SERS enhancement factor of 2 × 10 6 is evidence of their ability to perform as a SERS substrate. Although some iron oxide and gold substrates have shown comparable results, they typically rely on the use of Raman reporters or lengthy bioconjugation processes to target a biomolecule of interest. , Our bioseparation and bioidentification was performed rapidly (∼30 m) with only a mild reducing agent, TCEP, used to encourage binding, and our sensing was performed without the need for reporter molecules.…”

“…Although some iron oxide and gold substrates have shown comparable results, they typically rely on the use of Raman reporters or lengthy bioconjugation processes to target a biomolecule of interest. 51,52 Our bioseparation and bioidentification was performed rapidly (∼30 m) with only a mild reducing agent, TCEP, used to encourage binding, and our sensing was performed without the need for reporter molecules.…”

Recent Advances in Surface-Enhanced Raman Scattering Magnetic Plasmonic Particles for Bioapplications

“…Therefore, magnetic SERS has been applied in disease-based protein detection. Target proteins include human immunoglobulin G [ 20 , 36 ], prostate specific antigen [ 26 , 35 , 93 ], urinary erythropoietin [ 94 ], sepsis-specific biomarkers [ 95 ], carcinoembryonic antigen [ 96 ], thrombin, platelet derived growth factor BB, immunoglobulin E [ 62 ], and glycated hemoglobin [ 97 ]. These MPPs are designed with very specific components, such as antibodies, aptamers or chemical molecules [ 62 , 95 , 97 ].…”

“…These MPPs are designed with very specific components, such as antibodies, aptamers or chemical molecules [ 62 , 95 , 97 ]. The MPPs enable easy extraction, isolation, collection, and purification of the specific target from biological samples [ 62 , 93 , 94 , 95 , 96 , 97 ] and facilitate measurement at low concentrations.…”

Annual banned‐substance review: Analytical approaches in human sports drug testing

“…Further, urine was found to be the preferred matrix over plasma for targeting microdosed EPO in sports drug testing. An orthogonal approach towards detecting rhEPO in human urine was presented by Selbes et al, who suggested the use of a surface‐enhanced Raman probe for extraction and analysis of the target analyte . By means of the same anti‐EPO antibody clone AE7A5 as mentioned above, which was immobilized via a 11‐mercaptoundecanoic acid‐derived linker to a gold surface for extraction and a Raman label‐carrying rod‐shaped gold nanoparticle, a sensitive test method for EPO was presented.…”